#METABOLOMICS WORKBENCH CaoJing_20231121_191413 DATATRACK_ID:4472 STUDY_ID:ST002983 ANALYSIS_ID:AN004902 PROJECT_ID:PR001858 VERSION 1 CREATED_ON November 22, 2023, 6:35 pm #PROJECT PR:PROJECT_TITLE Deciphering the metabolic heterogeneity of hematopoietic stem cells with PR:PROJECT_TITLE single-cell resolution PR:PROJECT_TYPE Cell Metabolism PR:PROJECT_SUMMARY Metabolic status is crucial for stem cell functions; however, the metabolic PR:PROJECT_SUMMARY heterogeneity of endogenous stem cells has never been directly assessed. Here, PR:PROJECT_SUMMARY we develop a platform for high-throughput single-cell metabolomics (hi-scMet) of PR:PROJECT_SUMMARY hematopoietic stem cells (HSCs). By combining flow cytometric isolation and PR:PROJECT_SUMMARY nanoparticle-enhanced laser desorption/ionization mass spectrometry, we PR:PROJECT_SUMMARY routinely detected >100 features from single cells. We mapped the single-cell PR:PROJECT_SUMMARY metabolomes of all hematopoietic cell populations, and HSC subpopulations with PR:PROJECT_SUMMARY different division times, detecting 33 features whose levels exhibited trending PR:PROJECT_SUMMARY changes during HSC proliferation. We found progressive activation of oxidative PR:PROJECT_SUMMARY pentose phosphate pathway (OxiPPP) from dormant to active HSCs. Genetic or PR:PROJECT_SUMMARY pharmacological interference with OxiPPP increased reactive oxygen species level PR:PROJECT_SUMMARY in HSCs, reducing HSC self-renewal upon oxidative stress. Together, our work PR:PROJECT_SUMMARY uncovers the metabolic dynamics during HSC proliferation, reveals a role of PR:PROJECT_SUMMARY OxiPPP for HSC activation, and illustrates the utility of hi-scMet in dissecting PR:PROJECT_SUMMARY metabolic heterogeneity of immunophenotypically defined cell populations. PR:INSTITUTE Shanghai Jiao Tong University PR:LAST_NAME CAO PR:FIRST_NAME JING PR:ADDRESS 1954 Huashan Road, Shanghai, Shanghai, 200030, China PR:EMAIL caojing1@sjtu.edu.cn PR:PHONE 15201957271 PR:FUNDING_SOURCE This work was supported by the National Key R&D Program (2022YFE0103500, PR:FUNDING_SOURCE 2018YFA0107200, 2021YFF0703500, and 2022YFC2502800), Medical-Engineering Joint PR:FUNDING_SOURCE Funds of Shanghai Jiao Tong University (YG2021ZD09, YG2022QN107, YG2023ZD08), PR:FUNDING_SOURCE Haihe Laboratory of Cell Ecosystem Innovation Fund (22HHXBSS00016), the National PR:FUNDING_SOURCE Natural Science Foundation of China (32270785, 31771637, 81730006, 81971771, PR:FUNDING_SOURCE 22074044, 22122404) and the CAS Youth Interdisciplinary Team (JCTD-2021-12). PR:FUNDING_SOURCE This work was also sponsored by Shanghai Municipal Science and Technology Major PR:FUNDING_SOURCE Project Fund, Shanghai Science and Technology Commission (22XD1424000, PR:FUNDING_SOURCE 22ZR1469000), Shanghai Institutions of Higher Learning PR:FUNDING_SOURCE (2021-01-07-00-02-E00083), Innovative Research Team of High-Level Local PR:FUNDING_SOURCE Universities in Shanghai (SHSMU-ZDCX20210700), Innovation Group Project of PR:FUNDING_SOURCE Shanghai Municipal Health Comission (2019CXJQ03), Innovation Research Plan of PR:FUNDING_SOURCE Shanghai Municipal Education Commission (ZXWF082101), National Research Center PR:FUNDING_SOURCE for Translational Medicine Shanghai (TMSK-2021-124, NRCTM(SH)-2021-06), and PR:FUNDING_SOURCE Fundamental Research Funds for the Central Universities. #STUDY ST:STUDY_TITLE Deciphering the metabolic heterogeneity of hematopoietic stem cells with ST:STUDY_TITLE single-cell resolution ST:STUDY_SUMMARY Metabolic status is crucial for stem cell functions; however, the metabolic ST:STUDY_SUMMARY heterogeneity of endogenous stem cells has never been directly assessed. Here, ST:STUDY_SUMMARY we develop a platform for high-throughput single-cell metabolomics (hi-scMet) of ST:STUDY_SUMMARY hematopoietic stem cells (HSCs). By combining flow cytometric isolation and ST:STUDY_SUMMARY nanoparticle-enhanced laser desorption/ionization mass spectrometry, we ST:STUDY_SUMMARY routinely detected >100 features from single cells. We mapped the single-cell ST:STUDY_SUMMARY metabolomes of all hematopoietic cell populations, and HSC subpopulations with ST:STUDY_SUMMARY different division times, detecting 33 features whose levels exhibited trending ST:STUDY_SUMMARY changes during HSC proliferation. We found progressive activation of oxidative ST:STUDY_SUMMARY pentose phosphate pathway (OxiPPP) from dormant to active HSCs. Genetic or ST:STUDY_SUMMARY pharmacological interference with OxiPPP increased reactive oxygen species level ST:STUDY_SUMMARY in HSCs, reducing HSC self-renewal upon oxidative stress. Together, our work ST:STUDY_SUMMARY uncovers the metabolic dynamics during HSC proliferation, reveals a role of ST:STUDY_SUMMARY OxiPPP for HSC activation, and illustrates the utility of hi-scMet in dissecting ST:STUDY_SUMMARY metabolic heterogeneity of immunophenotypically defined cell populations. ST:INSTITUTE Shanghai Jiao Tong University ST:LAST_NAME CAO ST:FIRST_NAME JING ST:ADDRESS 1954 Huashan Road, Shanghai, Shanghai, 200030, China ST:EMAIL caojing1@sjtu.edu.cn ST:PHONE +8615201957271 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - HSCa_1 Celltype:HSCa RAW_FILE_NAME=HSCa_1_1.txt; RAW_FILE_NAME=HSCa_1_2.txt; RAW_FILE_NAME=HSCa_1_3.txt; RAW_FILE_NAME=HSCa_1_4.txt; RAW_FILE_NAME=HSCa_1_5.txt SUBJECT_SAMPLE_FACTORS - HSCa_2 Celltype:HSCa RAW_FILE_NAME=HSCa_2_1.txt; RAW_FILE_NAME=HSCa_2_2.txt; RAW_FILE_NAME=HSCa_2_3.txt; RAW_FILE_NAME=HSCa_2_4.txt; RAW_FILE_NAME=HSCa_2_5.txt SUBJECT_SAMPLE_FACTORS - HSCa_3 Celltype:HSCa RAW_FILE_NAME=HSCa_3_1.txt; RAW_FILE_NAME=HSCa_3_2.txt; RAW_FILE_NAME=HSCa_3_3.txt; RAW_FILE_NAME=HSCa_3_4.txt; RAW_FILE_NAME=HSCa_3_5.txt SUBJECT_SAMPLE_FACTORS - HSCa_4 Celltype:HSCa RAW_FILE_NAME=HSCa_4_1.txt; 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RAW_FILE_NAME=HSCb_14_4.txt; RAW_FILE_NAME=HSCb_14_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_15 Celltype:HSCb RAW_FILE_NAME=HSCb_15_1.txt; RAW_FILE_NAME=HSCb_15_2.txt; RAW_FILE_NAME=HSCb_15_3.txt; RAW_FILE_NAME=HSCb_15_4.txt; RAW_FILE_NAME=HSCb_15_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_16 Celltype:HSCb RAW_FILE_NAME=HSCb_16_1.txt; RAW_FILE_NAME=HSCb_16_2.txt; RAW_FILE_NAME=HSCb_16_3.txt; RAW_FILE_NAME=HSCb_16_4.txt; RAW_FILE_NAME=HSCb_16_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_17 Celltype:HSCb RAW_FILE_NAME=HSCb_17_1.txt; RAW_FILE_NAME=HSCb_17_2.txt; RAW_FILE_NAME=HSCb_17_3.txt; RAW_FILE_NAME=HSCb_17_4.txt; RAW_FILE_NAME=HSCb_17_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_18 Celltype:HSCb RAW_FILE_NAME=HSCb_18_1.txt; RAW_FILE_NAME=HSCb_18_2.txt; RAW_FILE_NAME=HSCb_18_3.txt; RAW_FILE_NAME=HSCb_18_4.txt; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - HSCb_19 Celltype:HSCb RAW_FILE_NAME=HSCb_19_1.txt; RAW_FILE_NAME=HSCb_19_2.txt; RAW_FILE_NAME=HSCb_19_3.txt; RAW_FILE_NAME=HSCb_19_4.txt; RAW_FILE_NAME=HSCb_19_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_20 Celltype:HSCb RAW_FILE_NAME=HSCb_20_1.txt; RAW_FILE_NAME=HSCb_20_2.txt; RAW_FILE_NAME=HSCb_20_3.txt; RAW_FILE_NAME=HSCb_20_4.txt; RAW_FILE_NAME=HSCb_20_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_21 Celltype:HSCb RAW_FILE_NAME=HSCb_21_1.txt; RAW_FILE_NAME=HSCb_21_2.txt; RAW_FILE_NAME=HSCb_21_3.txt; RAW_FILE_NAME=HSCb_21_4.txt; RAW_FILE_NAME=HSCb_21_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_22 Celltype:HSCb RAW_FILE_NAME=HSCb_22_1.txt; RAW_FILE_NAME=HSCb_22_2.txt; RAW_FILE_NAME=HSCb_22_3.txt; RAW_FILE_NAME=HSCb_22_4.txt; RAW_FILE_NAME=HSCb_22_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_23 Celltype:HSCb RAW_FILE_NAME=HSCb_23_1.txt; RAW_FILE_NAME=HSCb_23_2.txt; RAW_FILE_NAME=HSCb_23_3.txt; RAW_FILE_NAME=HSCb_23_4.txt; RAW_FILE_NAME=HSCb_23_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_24 Celltype:HSCb RAW_FILE_NAME=HSCb_24_1.txt; RAW_FILE_NAME=HSCb_24_2.txt; RAW_FILE_NAME=HSCb_24_3.txt; RAW_FILE_NAME=HSCb_24_4.txt; RAW_FILE_NAME=HSCb_24_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_25 Celltype:HSCb RAW_FILE_NAME=HSCb_25_1.txt; RAW_FILE_NAME=HSCb_25_2.txt; RAW_FILE_NAME=HSCb_25_3.txt; RAW_FILE_NAME=HSCb_25_4.txt; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - HSCb_26 Celltype:HSCb RAW_FILE_NAME=HSCb_26_1.txt; RAW_FILE_NAME=HSCb_26_2.txt; RAW_FILE_NAME=HSCb_26_3.txt; RAW_FILE_NAME=HSCb_26_4.txt; RAW_FILE_NAME=HSCb_26_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_27 Celltype:HSCb RAW_FILE_NAME=HSCb_27_1.txt; RAW_FILE_NAME=HSCb_27_2.txt; RAW_FILE_NAME=HSCb_27_3.txt; RAW_FILE_NAME=HSCb_27_4.txt; RAW_FILE_NAME=HSCb_27_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_28 Celltype:HSCb RAW_FILE_NAME=HSCb_28_1.txt; RAW_FILE_NAME=HSCb_28_2.txt; RAW_FILE_NAME=HSCb_28_3.txt; RAW_FILE_NAME=HSCb_28_4.txt; RAW_FILE_NAME=HSCb_28_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_29 Celltype:HSCb RAW_FILE_NAME=HSCb_29_1.txt; RAW_FILE_NAME=HSCb_29_2.txt; RAW_FILE_NAME=HSCb_29_3.txt; RAW_FILE_NAME=HSCb_29_4.txt; RAW_FILE_NAME=HSCb_29_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_30 Celltype:HSCb RAW_FILE_NAME=HSCb_30_1.txt; RAW_FILE_NAME=HSCb_30_2.txt; RAW_FILE_NAME=HSCb_30_3.txt; RAW_FILE_NAME=HSCb_30_4.txt; RAW_FILE_NAME=HSCb_30_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_31 Celltype:HSCb RAW_FILE_NAME=HSCb_31_1.txt; RAW_FILE_NAME=HSCb_31_2.txt; RAW_FILE_NAME=HSCb_31_3.txt; RAW_FILE_NAME=HSCb_31_4.txt; RAW_FILE_NAME=HSCb_31_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_32 Celltype:HSCb RAW_FILE_NAME=HSCb_32_1.txt; RAW_FILE_NAME=HSCb_32_2.txt; RAW_FILE_NAME=HSCb_32_3.txt; RAW_FILE_NAME=HSCb_32_4.txt; RAW_FILE_NAME=HSCb_32_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_33 Celltype:HSCb RAW_FILE_NAME=HSCb_33_1.txt; RAW_FILE_NAME=HSCb_33_2.txt; RAW_FILE_NAME=HSCb_33_3.txt; RAW_FILE_NAME=HSCb_33_4.txt; RAW_FILE_NAME=HSCb_33_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_34 Celltype:HSCb RAW_FILE_NAME=HSCb_34_1.txt; RAW_FILE_NAME=HSCb_34_2.txt; RAW_FILE_NAME=HSCb_34_3.txt; RAW_FILE_NAME=HSCb_34_4.txt; RAW_FILE_NAME=HSCb_34_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_35 Celltype:HSCb RAW_FILE_NAME=HSCb_35_1.txt; RAW_FILE_NAME=HSCb_35_2.txt; RAW_FILE_NAME=HSCb_35_3.txt; RAW_FILE_NAME=HSCb_35_4.txt; RAW_FILE_NAME=HSCb_35_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_36 Celltype:HSCb RAW_FILE_NAME=HSCb_36_1.txt; RAW_FILE_NAME=HSCb_36_2.txt; RAW_FILE_NAME=HSCb_36_3.txt; RAW_FILE_NAME=HSCb_36_4.txt; RAW_FILE_NAME=HSCb_36_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_37 Celltype:HSCb RAW_FILE_NAME=HSCb_37_1.txt; RAW_FILE_NAME=HSCb_37_2.txt; RAW_FILE_NAME=HSCb_37_3.txt; RAW_FILE_NAME=HSCb_37_4.txt; RAW_FILE_NAME=HSCb_37_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_38 Celltype:HSCb RAW_FILE_NAME=HSCb_38_1.txt; RAW_FILE_NAME=HSCb_38_2.txt; RAW_FILE_NAME=HSCb_38_3.txt; RAW_FILE_NAME=HSCb_38_4.txt; RAW_FILE_NAME=HSCb_38_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_39 Celltype:HSCb RAW_FILE_NAME=HSCb_39_1.txt; RAW_FILE_NAME=HSCb_39_2.txt; RAW_FILE_NAME=HSCb_39_3.txt; RAW_FILE_NAME=HSCb_39_4.txt; RAW_FILE_NAME=HSCb_39_5.txt SUBJECT_SAMPLE_FACTORS - HSCb_40 Celltype:HSCb RAW_FILE_NAME=HSCb_40_1.txt; RAW_FILE_NAME=HSCb_40_2.txt; RAW_FILE_NAME=HSCb_40_3.txt; RAW_FILE_NAME=HSCb_40_4.txt; RAW_FILE_NAME=HSCb_40_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_1 Celltype:HSCc RAW_FILE_NAME=HSCc_1_1.txt; RAW_FILE_NAME=HSCc_1_2.txt; RAW_FILE_NAME=HSCc_1_3.txt; RAW_FILE_NAME=HSCc_1_4.txt; RAW_FILE_NAME=HSCc_1_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_2 Celltype:HSCc RAW_FILE_NAME=HSCc_2_1.txt; RAW_FILE_NAME=HSCc_2_2.txt; RAW_FILE_NAME=HSCc_2_3.txt; RAW_FILE_NAME=HSCc_2_4.txt; RAW_FILE_NAME=HSCc_2_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_3 Celltype:HSCc RAW_FILE_NAME=HSCc_3_1.txt; RAW_FILE_NAME=HSCc_3_2.txt; RAW_FILE_NAME=HSCc_3_3.txt; RAW_FILE_NAME=HSCc_3_4.txt; RAW_FILE_NAME=HSCc_3_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_4 Celltype:HSCc RAW_FILE_NAME=HSCc_4_1.txt; RAW_FILE_NAME=HSCc_4_2.txt; RAW_FILE_NAME=HSCc_4_3.txt; RAW_FILE_NAME=HSCc_4_4.txt; RAW_FILE_NAME=HSCc_4_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_5 Celltype:HSCc RAW_FILE_NAME=HSCc_5_1.txt; RAW_FILE_NAME=HSCc_5_2.txt; RAW_FILE_NAME=HSCc_5_3.txt; RAW_FILE_NAME=HSCc_5_4.txt; RAW_FILE_NAME=HSCc_5_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_6 Celltype:HSCc RAW_FILE_NAME=HSCc_6_1.txt; RAW_FILE_NAME=HSCc_6_2.txt; RAW_FILE_NAME=HSCc_6_3.txt; RAW_FILE_NAME=HSCc_6_4.txt; RAW_FILE_NAME=HSCc_6_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_7 Celltype:HSCc RAW_FILE_NAME=HSCc_7_1.txt; RAW_FILE_NAME=HSCc_7_2.txt; RAW_FILE_NAME=HSCc_7_3.txt; RAW_FILE_NAME=HSCc_7_4.txt; RAW_FILE_NAME=HSCc_7_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_8 Celltype:HSCc RAW_FILE_NAME=HSCc_8_1.txt; RAW_FILE_NAME=HSCc_8_2.txt; RAW_FILE_NAME=HSCc_8_3.txt; RAW_FILE_NAME=HSCc_8_4.txt; RAW_FILE_NAME=HSCc_8_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_9 Celltype:HSCc RAW_FILE_NAME=HSCc_9_1.txt; RAW_FILE_NAME=HSCc_9_2.txt; RAW_FILE_NAME=HSCc_9_3.txt; RAW_FILE_NAME=HSCc_9_4.txt; RAW_FILE_NAME=HSCc_9_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_10 Celltype:HSCc RAW_FILE_NAME=HSCc_10_1.txt; RAW_FILE_NAME=HSCc_10_2.txt; RAW_FILE_NAME=HSCc_10_3.txt; RAW_FILE_NAME=HSCc_10_4.txt; RAW_FILE_NAME=HSCc_10_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_11 Celltype:HSCc RAW_FILE_NAME=HSCc_11_1.txt; RAW_FILE_NAME=HSCc_11_2.txt; RAW_FILE_NAME=HSCc_11_3.txt; RAW_FILE_NAME=HSCc_11_4.txt; RAW_FILE_NAME=HSCc_11_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_12 Celltype:HSCc RAW_FILE_NAME=HSCc_12_1.txt; RAW_FILE_NAME=HSCc_12_2.txt; RAW_FILE_NAME=HSCc_12_3.txt; RAW_FILE_NAME=HSCc_12_4.txt; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - HSCc_13 Celltype:HSCc RAW_FILE_NAME=HSCc_13_1.txt; RAW_FILE_NAME=HSCc_13_2.txt; RAW_FILE_NAME=HSCc_13_3.txt; RAW_FILE_NAME=HSCc_13_4.txt; RAW_FILE_NAME=HSCc_13_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_14 Celltype:HSCc RAW_FILE_NAME=HSCc_14_1.txt; RAW_FILE_NAME=HSCc_14_2.txt; RAW_FILE_NAME=HSCc_14_3.txt; RAW_FILE_NAME=HSCc_14_4.txt; RAW_FILE_NAME=HSCc_14_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_15 Celltype:HSCc RAW_FILE_NAME=HSCc_15_1.txt; RAW_FILE_NAME=HSCc_15_2.txt; RAW_FILE_NAME=HSCc_15_3.txt; RAW_FILE_NAME=HSCc_15_4.txt; RAW_FILE_NAME=HSCc_15_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_16 Celltype:HSCc RAW_FILE_NAME=HSCc_16_1.txt; RAW_FILE_NAME=HSCc_16_2.txt; RAW_FILE_NAME=HSCc_16_3.txt; RAW_FILE_NAME=HSCc_16_4.txt; RAW_FILE_NAME=HSCc_16_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_17 Celltype:HSCc RAW_FILE_NAME=HSCc_17_1.txt; RAW_FILE_NAME=HSCc_17_2.txt; RAW_FILE_NAME=HSCc_17_3.txt; RAW_FILE_NAME=HSCc_17_4.txt; RAW_FILE_NAME=HSCc_17_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_18 Celltype:HSCc RAW_FILE_NAME=HSCc_18_1.txt; RAW_FILE_NAME=HSCc_18_2.txt; RAW_FILE_NAME=HSCc_18_3.txt; RAW_FILE_NAME=HSCc_18_4.txt; RAW_FILE_NAME=HSCc_18_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_19 Celltype:HSCc RAW_FILE_NAME=HSCc_19_1.txt; RAW_FILE_NAME=HSCc_19_2.txt; RAW_FILE_NAME=HSCc_19_3.txt; RAW_FILE_NAME=HSCc_19_4.txt; RAW_FILE_NAME=HSCc_19_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_20 Celltype:HSCc RAW_FILE_NAME=HSCc_20_1.txt; RAW_FILE_NAME=HSCc_20_2.txt; RAW_FILE_NAME=HSCc_20_3.txt; RAW_FILE_NAME=HSCc_20_4.txt; RAW_FILE_NAME=HSCc_20_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_21 Celltype:HSCc RAW_FILE_NAME=HSCc_21_1.txt; RAW_FILE_NAME=HSCc_21_2.txt; RAW_FILE_NAME=HSCc_21_3.txt; RAW_FILE_NAME=HSCc_21_4.txt; RAW_FILE_NAME=HSCc_21_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_22 Celltype:HSCc RAW_FILE_NAME=HSCc_22_1.txt; RAW_FILE_NAME=HSCc_22_2.txt; RAW_FILE_NAME=HSCc_22_3.txt; RAW_FILE_NAME=HSCc_22_4.txt; RAW_FILE_NAME=HSCc_22_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_23 Celltype:HSCc RAW_FILE_NAME=HSCc_23_1.txt; RAW_FILE_NAME=HSCc_23_2.txt; RAW_FILE_NAME=HSCc_23_3.txt; RAW_FILE_NAME=HSCc_23_4.txt; RAW_FILE_NAME=HSCc_23_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_24 Celltype:HSCc RAW_FILE_NAME=HSCc_24_1.txt; RAW_FILE_NAME=HSCc_24_2.txt; RAW_FILE_NAME=HSCc_24_3.txt; RAW_FILE_NAME=HSCc_24_4.txt; RAW_FILE_NAME=HSCc_24_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_25 Celltype:HSCc RAW_FILE_NAME=HSCc_25_1.txt; RAW_FILE_NAME=HSCc_25_2.txt; RAW_FILE_NAME=HSCc_25_3.txt; RAW_FILE_NAME=HSCc_25_4.txt; RAW_FILE_NAME=HSCc_25_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_26 Celltype:HSCc RAW_FILE_NAME=HSCc_26_1.txt; RAW_FILE_NAME=HSCc_26_2.txt; RAW_FILE_NAME=HSCc_26_3.txt; RAW_FILE_NAME=HSCc_26_4.txt; RAW_FILE_NAME=HSCc_26_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_27 Celltype:HSCc RAW_FILE_NAME=HSCc_27_1.txt; RAW_FILE_NAME=HSCc_27_2.txt; RAW_FILE_NAME=HSCc_27_3.txt; RAW_FILE_NAME=HSCc_27_4.txt; RAW_FILE_NAME=HSCc_27_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_28 Celltype:HSCc RAW_FILE_NAME=HSCc_28_1.txt; RAW_FILE_NAME=HSCc_28_2.txt; RAW_FILE_NAME=HSCc_28_3.txt; RAW_FILE_NAME=HSCc_28_4.txt; RAW_FILE_NAME=HSCc_28_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_29 Celltype:HSCc RAW_FILE_NAME=HSCc_29_1.txt; RAW_FILE_NAME=HSCc_29_2.txt; RAW_FILE_NAME=HSCc_29_3.txt; RAW_FILE_NAME=HSCc_29_4.txt; RAW_FILE_NAME=HSCc_29_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_30 Celltype:HSCc RAW_FILE_NAME=HSCc_30_1.txt; RAW_FILE_NAME=HSCc_30_2.txt; RAW_FILE_NAME=HSCc_30_3.txt; RAW_FILE_NAME=HSCc_30_4.txt; RAW_FILE_NAME=HSCc_30_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_31 Celltype:HSCc RAW_FILE_NAME=HSCc_31_1.txt; RAW_FILE_NAME=HSCc_31_2.txt; RAW_FILE_NAME=HSCc_31_3.txt; RAW_FILE_NAME=HSCc_31_4.txt; RAW_FILE_NAME=HSCc_31_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_32 Celltype:HSCc RAW_FILE_NAME=HSCc_32_1.txt; RAW_FILE_NAME=HSCc_32_2.txt; RAW_FILE_NAME=HSCc_32_3.txt; RAW_FILE_NAME=HSCc_32_4.txt; RAW_FILE_NAME=HSCc_32_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_33 Celltype:HSCc RAW_FILE_NAME=HSCc_33_1.txt; RAW_FILE_NAME=HSCc_33_2.txt; RAW_FILE_NAME=HSCc_33_3.txt; RAW_FILE_NAME=HSCc_33_4.txt; RAW_FILE_NAME=HSCc_33_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_34 Celltype:HSCc RAW_FILE_NAME=HSCc_34_1.txt; RAW_FILE_NAME=HSCc_34_2.txt; RAW_FILE_NAME=HSCc_34_3.txt; RAW_FILE_NAME=HSCc_34_4.txt; RAW_FILE_NAME=HSCc_34_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_35 Celltype:HSCc RAW_FILE_NAME=HSCc_35_1.txt; RAW_FILE_NAME=HSCc_35_2.txt; RAW_FILE_NAME=HSCc_35_3.txt; RAW_FILE_NAME=HSCc_35_4.txt; RAW_FILE_NAME=HSCc_35_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_36 Celltype:HSCc RAW_FILE_NAME=HSCc_36_1.txt; RAW_FILE_NAME=HSCc_36_2.txt; RAW_FILE_NAME=HSCc_36_3.txt; RAW_FILE_NAME=HSCc_36_4.txt; RAW_FILE_NAME=HSCc_36_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_37 Celltype:HSCc RAW_FILE_NAME=HSCc_37_1.txt; RAW_FILE_NAME=HSCc_37_2.txt; RAW_FILE_NAME=HSCc_37_3.txt; RAW_FILE_NAME=HSCc_37_4.txt; RAW_FILE_NAME=HSCc_37_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_38 Celltype:HSCc RAW_FILE_NAME=HSCc_38_1.txt; RAW_FILE_NAME=HSCc_38_2.txt; RAW_FILE_NAME=HSCc_38_3.txt; RAW_FILE_NAME=HSCc_38_4.txt; RAW_FILE_NAME=HSCc_38_5.txt SUBJECT_SAMPLE_FACTORS - HSCc_39 Celltype:HSCc RAW_FILE_NAME=HSCc_39_1.txt; RAW_FILE_NAME=HSCc_39_2.txt; RAW_FILE_NAME=HSCc_39_3.txt; RAW_FILE_NAME=HSCc_39_4.txt; RAW_FILE_NAME=HSCc_39_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_1 Celltype:HSCd RAW_FILE_NAME=HSCd_1_1.txt; RAW_FILE_NAME=HSCd_1_2.txt; RAW_FILE_NAME=HSCd_1_3.txt; RAW_FILE_NAME=HSCd_1_4.txt; RAW_FILE_NAME=HSCd_1_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_2 Celltype:HSCd RAW_FILE_NAME=HSCd_2_1.txt; RAW_FILE_NAME=HSCd_2_2.txt; RAW_FILE_NAME=HSCd_2_3.txt; RAW_FILE_NAME=HSCd_2_4.txt; RAW_FILE_NAME=HSCd_2_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_3 Celltype:HSCd RAW_FILE_NAME=HSCd_3_1.txt; RAW_FILE_NAME=HSCd_3_2.txt; RAW_FILE_NAME=HSCd_3_3.txt; RAW_FILE_NAME=HSCd_3_4.txt; RAW_FILE_NAME=HSCd_3_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_4 Celltype:HSCd RAW_FILE_NAME=HSCd_4_1.txt; RAW_FILE_NAME=HSCd_4_2.txt; RAW_FILE_NAME=HSCd_4_3.txt; RAW_FILE_NAME=HSCd_4_4.txt; RAW_FILE_NAME=HSCd_4_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_5 Celltype:HSCd RAW_FILE_NAME=HSCd_5_1.txt; RAW_FILE_NAME=HSCd_5_2.txt; RAW_FILE_NAME=HSCd_5_3.txt; RAW_FILE_NAME=HSCd_5_4.txt; RAW_FILE_NAME=HSCd_5_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_6 Celltype:HSCd RAW_FILE_NAME=HSCd_6_1.txt; RAW_FILE_NAME=HSCd_6_2.txt; RAW_FILE_NAME=HSCd_6_3.txt; RAW_FILE_NAME=HSCd_6_4.txt; RAW_FILE_NAME=HSCd_6_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_7 Celltype:HSCd RAW_FILE_NAME=HSCd_7_1.txt; RAW_FILE_NAME=HSCd_7_2.txt; RAW_FILE_NAME=HSCd_7_3.txt; RAW_FILE_NAME=HSCd_7_4.txt; RAW_FILE_NAME=HSCd_7_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_8 Celltype:HSCd RAW_FILE_NAME=HSCd_8_1.txt; RAW_FILE_NAME=HSCd_8_2.txt; RAW_FILE_NAME=HSCd_8_3.txt; RAW_FILE_NAME=HSCd_8_4.txt; RAW_FILE_NAME=HSCd_8_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_9 Celltype:HSCd RAW_FILE_NAME=HSCd_9_1.txt; RAW_FILE_NAME=HSCd_9_2.txt; RAW_FILE_NAME=HSCd_9_3.txt; RAW_FILE_NAME=HSCd_9_4.txt; RAW_FILE_NAME=HSCd_9_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_10 Celltype:HSCd RAW_FILE_NAME=HSCd_10_1.txt; RAW_FILE_NAME=HSCd_10_2.txt; RAW_FILE_NAME=HSCd_10_3.txt; RAW_FILE_NAME=HSCd_10_4.txt; RAW_FILE_NAME=HSCd_10_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_11 Celltype:HSCd RAW_FILE_NAME=HSCd_11_1.txt; RAW_FILE_NAME=HSCd_11_2.txt; RAW_FILE_NAME=HSCd_11_3.txt; RAW_FILE_NAME=HSCd_11_4.txt; RAW_FILE_NAME=HSCd_11_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_12 Celltype:HSCd RAW_FILE_NAME=HSCd_12_1.txt; RAW_FILE_NAME=HSCd_12_2.txt; RAW_FILE_NAME=HSCd_12_3.txt; RAW_FILE_NAME=HSCd_12_4.txt; RAW_FILE_NAME=HSCd_12_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_13 Celltype:HSCd RAW_FILE_NAME=HSCd_13_1.txt; RAW_FILE_NAME=HSCd_13_2.txt; RAW_FILE_NAME=HSCd_13_3.txt; RAW_FILE_NAME=HSCd_13_4.txt; RAW_FILE_NAME=HSCd_13_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_14 Celltype:HSCd RAW_FILE_NAME=HSCd_14_1.txt; RAW_FILE_NAME=HSCd_14_2.txt; RAW_FILE_NAME=HSCd_14_3.txt; RAW_FILE_NAME=HSCd_14_4.txt; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - HSCd_15 Celltype:HSCd RAW_FILE_NAME=HSCd_15_1.txt; RAW_FILE_NAME=HSCd_15_2.txt; RAW_FILE_NAME=HSCd_15_3.txt; RAW_FILE_NAME=HSCd_15_4.txt; RAW_FILE_NAME=HSCd_15_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_16 Celltype:HSCd RAW_FILE_NAME=HSCd_16_1.txt; RAW_FILE_NAME=HSCd_16_2.txt; RAW_FILE_NAME=HSCd_16_3.txt; RAW_FILE_NAME=HSCd_16_4.txt; RAW_FILE_NAME=HSCd_16_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_17 Celltype:HSCd RAW_FILE_NAME=HSCd_17_1.txt; RAW_FILE_NAME=HSCd_17_2.txt; RAW_FILE_NAME=HSCd_17_3.txt; RAW_FILE_NAME=HSCd_17_4.txt; RAW_FILE_NAME=HSCd_17_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_18 Celltype:HSCd RAW_FILE_NAME=HSCd_18_1.txt; RAW_FILE_NAME=HSCd_18_2.txt; RAW_FILE_NAME=HSCd_18_3.txt; RAW_FILE_NAME=HSCd_18_4.txt; RAW_FILE_NAME=HSCd_18_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_19 Celltype:HSCd RAW_FILE_NAME=HSCd_19_1.txt; RAW_FILE_NAME=HSCd_19_2.txt; RAW_FILE_NAME=HSCd_19_3.txt; RAW_FILE_NAME=HSCd_19_4.txt; RAW_FILE_NAME=HSCd_19_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_20 Celltype:HSCd RAW_FILE_NAME=HSCd_20_1.txt; RAW_FILE_NAME=HSCd_20_2.txt; RAW_FILE_NAME=HSCd_20_3.txt; RAW_FILE_NAME=HSCd_20_4.txt; RAW_FILE_NAME=HSCd_20_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_21 Celltype:HSCd RAW_FILE_NAME=HSCd_21_1.txt; RAW_FILE_NAME=HSCd_21_2.txt; RAW_FILE_NAME=HSCd_21_3.txt; RAW_FILE_NAME=HSCd_21_4.txt; RAW_FILE_NAME=HSCd_21_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_22 Celltype:HSCd RAW_FILE_NAME=HSCd_22_1.txt; RAW_FILE_NAME=HSCd_22_2.txt; RAW_FILE_NAME=HSCd_22_3.txt; RAW_FILE_NAME=HSCd_22_4.txt; RAW_FILE_NAME=HSCd_22_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_23 Celltype:HSCd RAW_FILE_NAME=HSCd_23_1.txt; RAW_FILE_NAME=HSCd_23_2.txt; RAW_FILE_NAME=HSCd_23_3.txt; RAW_FILE_NAME=HSCd_23_4.txt; RAW_FILE_NAME=HSCd_23_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_24 Celltype:HSCd RAW_FILE_NAME=HSCd_24_1.txt; RAW_FILE_NAME=HSCd_24_2.txt; RAW_FILE_NAME=HSCd_24_3.txt; RAW_FILE_NAME=HSCd_24_4.txt; RAW_FILE_NAME=HSCd_24_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_25 Celltype:HSCd RAW_FILE_NAME=HSCd_25_1.txt; RAW_FILE_NAME=HSCd_25_2.txt; RAW_FILE_NAME=HSCd_25_3.txt; RAW_FILE_NAME=HSCd_25_4.txt; RAW_FILE_NAME=HSCd_25_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_26 Celltype:HSCd RAW_FILE_NAME=HSCd_26_1.txt; RAW_FILE_NAME=HSCd_26_2.txt; RAW_FILE_NAME=HSCd_26_3.txt; RAW_FILE_NAME=HSCd_26_4.txt; RAW_FILE_NAME=HSCd_26_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_27 Celltype:HSCd RAW_FILE_NAME=HSCd_27_1.txt; RAW_FILE_NAME=HSCd_27_2.txt; RAW_FILE_NAME=HSCd_27_3.txt; RAW_FILE_NAME=HSCd_27_4.txt; RAW_FILE_NAME=HSCd_27_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_28 Celltype:HSCd RAW_FILE_NAME=HSCd_28_1.txt; RAW_FILE_NAME=HSCd_28_2.txt; RAW_FILE_NAME=HSCd_28_3.txt; RAW_FILE_NAME=HSCd_28_4.txt; RAW_FILE_NAME=HSCd_28_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_29 Celltype:HSCd RAW_FILE_NAME=HSCd_29_1.txt; RAW_FILE_NAME=HSCd_29_2.txt; RAW_FILE_NAME=HSCd_29_3.txt; RAW_FILE_NAME=HSCd_29_4.txt; RAW_FILE_NAME=HSCd_29_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_30 Celltype:HSCd RAW_FILE_NAME=HSCd_30_1.txt; RAW_FILE_NAME=HSCd_30_2.txt; RAW_FILE_NAME=HSCd_30_3.txt; RAW_FILE_NAME=HSCd_30_4.txt; RAW_FILE_NAME=HSCd_30_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_31 Celltype:HSCd RAW_FILE_NAME=HSCd_31_1.txt; RAW_FILE_NAME=HSCd_31_2.txt; RAW_FILE_NAME=HSCd_31_3.txt; RAW_FILE_NAME=HSCd_31_4.txt; RAW_FILE_NAME=HSCd_31_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_32 Celltype:HSCd RAW_FILE_NAME=HSCd_32_1.txt; RAW_FILE_NAME=HSCd_32_2.txt; RAW_FILE_NAME=HSCd_32_3.txt; RAW_FILE_NAME=HSCd_32_4.txt; RAW_FILE_NAME=HSCd_32_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_33 Celltype:HSCd RAW_FILE_NAME=HSCd_33_1.txt; RAW_FILE_NAME=HSCd_33_2.txt; RAW_FILE_NAME=HSCd_33_3.txt; RAW_FILE_NAME=HSCd_33_4.txt; RAW_FILE_NAME=HSCd_33_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_34 Celltype:HSCd RAW_FILE_NAME=HSCd_34_1.txt; RAW_FILE_NAME=HSCd_34_2.txt; RAW_FILE_NAME=HSCd_34_3.txt; RAW_FILE_NAME=HSCd_34_4.txt; RAW_FILE_NAME=HSCd_34_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_35 Celltype:HSCd RAW_FILE_NAME=HSCd_35_1.txt; RAW_FILE_NAME=HSCd_35_2.txt; RAW_FILE_NAME=HSCd_35_3.txt; RAW_FILE_NAME=HSCd_35_4.txt; RAW_FILE_NAME=HSCd_35_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_36 Celltype:HSCd RAW_FILE_NAME=HSCd_36_1.txt; RAW_FILE_NAME=HSCd_36_2.txt; RAW_FILE_NAME=HSCd_36_3.txt; RAW_FILE_NAME=HSCd_36_4.txt; RAW_FILE_NAME=HSCd_36_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_37 Celltype:HSCd RAW_FILE_NAME=HSCd_37_1.txt; RAW_FILE_NAME=HSCd_37_2.txt; RAW_FILE_NAME=HSCd_37_3.txt; RAW_FILE_NAME=HSCd_37_4.txt; RAW_FILE_NAME=HSCd_37_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_38 Celltype:HSCd RAW_FILE_NAME=HSCd_38_1.txt; RAW_FILE_NAME=HSCd_38_2.txt; RAW_FILE_NAME=HSCd_38_3.txt; RAW_FILE_NAME=HSCd_38_4.txt; RAW_FILE_NAME=HSCd_38_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_39 Celltype:HSCd RAW_FILE_NAME=HSCd_39_1.txt; RAW_FILE_NAME=HSCd_39_2.txt; RAW_FILE_NAME=HSCd_39_3.txt; RAW_FILE_NAME=HSCd_39_4.txt; RAW_FILE_NAME=HSCd_39_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_40 Celltype:HSCd RAW_FILE_NAME=HSCd_40_1.txt; RAW_FILE_NAME=HSCd_40_2.txt; RAW_FILE_NAME=HSCd_40_3.txt; RAW_FILE_NAME=HSCd_40_4.txt; RAW_FILE_NAME=HSCd_40_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_41 Celltype:HSCd RAW_FILE_NAME=HSCd_41_1.txt; RAW_FILE_NAME=HSCd_41_2.txt; RAW_FILE_NAME=HSCd_41_3.txt; RAW_FILE_NAME=HSCd_41_4.txt; RAW_FILE_NAME=HSCd_41_5.txt SUBJECT_SAMPLE_FACTORS - HSCd_42 Celltype:HSCd RAW_FILE_NAME=HSCd_42_1.txt; RAW_FILE_NAME=HSCd_42_2.txt; RAW_FILE_NAME=HSCd_42_3.txt; RAW_FILE_NAME=HSCd_42_4.txt; RAW_FILE_NAME=HSCd_42_5.txt #COLLECTION CO:COLLECTION_SUMMARY Bones were rapidly dissected and stored on ice in Ca2+- and Mg2+-free HBSS CO:COLLECTION_SUMMARY (ThermoFisher) plus 2% heat-inactivated fetal bovine serum (Gibco). Bone marrow CO:COLLECTION_SUMMARY cells were flushed out from the bone and then dissociated to single-cell CO:COLLECTION_SUMMARY suspension by gently passing through the needle then filtering through a 70-μm CO:COLLECTION_SUMMARY nylon mesh. The following antibodies were used to isolate hematopoietic cells: CO:COLLECTION_SUMMARY TER-119-FITC, CD3-FITC, CD5-FITC, CD8-FITC, B220-FITC, Gr-1-FITC, CO:COLLECTION_SUMMARY TER-119-APC780, CD3e-APC780, CD5-APC780, CD8-APC780, B220-APC780, Gr-1-APC780, CO:COLLECTION_SUMMARY TER-119-biotin, CD3-biotin, B220-biotin, Gr-1-biotin, c-kit-biotin, CO:COLLECTION_SUMMARY TER-119-PerCP/Cy5.5, Sca-1-PerCP/Cy5.5, MAC-1-APC, CD48-APC, CD16/32-APC, CO:COLLECTION_SUMMARY CD135-APC, CD127-PE, CD34-PE and CD3-PE, CD150-PE. FITC streptavidin, APC/Cy7 CO:COLLECTION_SUMMARY streptavidin and/or APC-R700 streptavidin were used for biotin-labeled CO:COLLECTION_SUMMARY antibodies. All reagents were acquired from BD Biosciences, eBiosciences or CO:COLLECTION_SUMMARY BioLegend. For isolation of HSPCs, cells were incubated with c-kit-biotin and CO:COLLECTION_SUMMARY paramagnetic microbeads, and then passed through an autoMACS magnetic separator. CO:COLLECTION_SUMMARY For isolation of CLPs, lineage was stained with Ter119-biotin, CD3-biotin, CO:COLLECTION_SUMMARY B220-biotin and Gr-1-biotin and paramagnetic microbeads. Then an autoMACS CO:COLLECTION_SUMMARY magnetic separator was used to enrich lineage- populations. To minimize CO:COLLECTION_SUMMARY metabolic changes, antibody-stained cells were fixed with 2% PFA (Sigma) for 15 CO:COLLECTION_SUMMARY minutes on ice. Flow cytometric analysis was performed with a BD LSRFortessa CO:COLLECTION_SUMMARY cytometer. To measure ROS levels of HSCs, antibody-stained cells were incubated CO:COLLECTION_SUMMARY with 5μm DCFDA for 15 minutes at 37°C before flow cytometric analysis. For CO:COLLECTION_SUMMARY metabolic detection, cells were sorted with a FACSAria SORP cytometer into CO:COLLECTION_SUMMARY 384-well plates containing 2.5-μl 80% methanol (Sigma) per well, and then CO:COLLECTION_SUMMARY centrifuged at 1500g for 5 mins at 4°C. Plates were sunk in liquid nitrogen for CO:COLLECTION_SUMMARY 10 mins to lyse cells and kept at -80°C before MS analysis. CO:SAMPLE_TYPE Stem cells #TREATMENT TR:TREATMENT_SUMMARY The following antibodies were used to isolate hematopoietic cells: TER-119-FITC, TR:TREATMENT_SUMMARY CD3-FITC, CD5-FITC, CD8-FITC, B220-FITC, Gr-1-FITC, TER-119-APC780, CD3e-APC780, TR:TREATMENT_SUMMARY CD5-APC780, CD8-APC780, B220-APC780, Gr-1-APC780, TER-119-biotin, CD3-biotin, TR:TREATMENT_SUMMARY B220-biotin, Gr-1-biotin, c-kit-biotin, TER-119-PerCP/Cy5.5, Sca-1-PerCP/Cy5.5, TR:TREATMENT_SUMMARY MAC-1-APC, CD48-APC, CD16/32-APC, CD135-APC, CD127-PE, CD34-PE and CD3-PE, TR:TREATMENT_SUMMARY CD150-PE. FITC streptavidin, APC/Cy7 streptavidin and/or APC-R700 streptavidin TR:TREATMENT_SUMMARY were used for biotin-labeled antibodies. All reagents were acquired from BD TR:TREATMENT_SUMMARY Biosciences, eBiosciences or BioLegend. For isolation of HSPCs, cells were TR:TREATMENT_SUMMARY incubated with c-kit-biotin and paramagnetic microbeads, and then passed through TR:TREATMENT_SUMMARY an autoMACS magnetic separator. For isolation of CLPs, lineage was stained with TR:TREATMENT_SUMMARY Ter119-biotin, CD3-biotin, B220-biotin and Gr-1-biotin and paramagnetic TR:TREATMENT_SUMMARY microbeads. Then an autoMACS magnetic separator was used to enrich lineage- TR:TREATMENT_SUMMARY populations. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For metabolic detection, cells were sorted with a FACSAria SORP cytometer into SP:SAMPLEPREP_SUMMARY 384-well plates containing 2.5-μl 80% methanol (Sigma) per well, and then SP:SAMPLEPREP_SUMMARY centrifuged at 1500g for 5 mins at 4°C. Plates were sunk in liquid nitrogen for SP:SAMPLEPREP_SUMMARY 10 mins to lyse cells and kept at -80°C before MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY None CH:CHROMATOGRAPHY_TYPE None (Direct infusion) CH:INSTRUMENT_NAME none CH:COLUMN_NAME none CH:SOLVENT_A none CH:SOLVENT_B none CH:FLOW_GRADIENT none CH:FLOW_RATE none CH:COLUMN_TEMPERATURE none #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Bruker Autoflex MALDI-TOF (/TOF)-MS MS:INSTRUMENT_TYPE MALDI-TOF-MS MS:MS_TYPE MALDI MS:ION_MODE POSITIVE MS:MS_COMMENTS MS acquisition Comments: MS experiments were conducted on the Autoflex MALDI-TOF MS:MS_COMMENTS (/TOF)-MS (Bruker Autoflex Speed) with Nd:YAG lasers (355 nm) and smart beam MS:MS_COMMENTS system. All MS experiments were performed in the reflector positive ion mode, MS:MS_COMMENTS with 2000 laser shots per analysis and 5 independent experiments per sample. MS:MS_COMMENTS Data processing Comments:MS data preprocessing was carried out on python version MS:MS_COMMENTS 3.8, including peak detection, peak filtration, standardization, and batch MS:MS_COMMENTS effect removal by ComBat algorithm. The heatmap, hierarchical clustering MS:MS_COMMENTS analysis, Pearson correlation analysis and PCA analysis were performed on MS:MS_COMMENTS MetaboAnalyst 5.0 (https://www.metaboanalyst.ca/). Machine learning and t-SNE MS:MS_COMMENTS clustering were conducted on Orange (version 3.25.0, the Bioinformatics Lab at MS:MS_COMMENTS University of Ljubljana, Slovenia). Software/procedures used for feature MS:MS_COMMENTS assignments: Python MS:MS_RESULTS_FILE ST002983_AN004902_Results.txt UNITS:intensity Has m/z:Yes Has RT:No RT units:No RT data #END