Summary of Study ST002114

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001192. The data can be accessed directly via it's Project DOI: 10.21228/M8MX3X This work is supported by NIH grant, U2C- DK119886.

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Study IDST002114
Study TitleInvolvement of Mieap in Cardiolipin metabolism (part I-revised)
Study SummaryCardiolipin (CL) alterations cause mitochondrial dysfunction. Mieap is involved in mitochondrial quality control (MQC). To investigate whether Mieap functions in MQC via regulation of CL metabolism, quantitative assessment of total CL and comparison of CL species conducted with A549 (Ad-Mieap infected vs. non-infected) and LS174T cells (LS174T-cont vs. Mieap-KD). The A549 cells were harvested 24 hr after infection with Ad-Mieap and were compared with non-infected cells by mass spectrometric analysis. The LS174T-cont and Mieap-KD cells incubated under a normal condition were harvested and subjected to mass spectrometric analysis.
Institute
National Cancer Center Japan Research Institute
Last NameIkari
First NameNaoki
Address5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan
Emailnikari@ncc.go.jp
Phone+81335422511
Submit Date2022-02-13
Raw Data AvailableYes
Raw Data File Type(s)cdf, raw(Thermo)
Analysis Type DetailLC-MS
Release Date2022-03-21
Release Version2
Release CommentsReplacement for ST001893 (datatrack 2779)
Naoki Ikari Naoki Ikari
https://dx.doi.org/10.21228/M8MX3X
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001192
Project DOI:doi: 10.21228/M8MX3X
Project Title:Mass Spectrometric Data of Cardiolipin in Cells under Mieap Expression
Project Summary:Mass spectrometric data of cardiolipin in A549 (Ad-Mieap infected vs. non-infected) and LS174T cells (LS174T-cont vs. Mieap-KD)
Institute:National Cancer Center Japan Research Institute
Last Name:Ikari
First Name:Naoki
Address:5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan
Email:nikari@ncc.go.jp
Phone:+81-3-3542-2511

Subject:

Subject ID:SU002199
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Cell Biosource Or Supplier:American Type Culture Collection
Cell Strain Details:A549 (tissue, lung cancer; gender, male), LS174T (tissue, colon cancer; gender, female)
Cell Counts:10,000,000

Factors:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Genotype/Treatment
SA202952A549 Ad-Mieap 4Ad-Mieap infection
SA202953A549 Ad-Mieap 1Ad-Mieap infection
SA202954A549 Ad-Mieap 3Ad-Mieap infection
SA202955A549 Ad-Mieap 2Ad-Mieap infection
SA202956LS174T Control 3Control
SA202957LS174T Control 4Control
SA202958LS174T Control 6Control
SA202959LS174T Control 5Control
SA202960LS174T Control 2Control
SA202961LS174T Control 1Control
SA202962LS174T Mieap-KD 5Mieap-knockdown
SA202963LS174T Mieap-KD 6Mieap-knockdown
SA202964LS174T Mieap-KD 1Mieap-knockdown
SA202965LS174T Mieap-KD 4Mieap-knockdown
SA202966LS174T Mieap-KD 2Mieap-knockdown
SA202967LS174T Mieap-KD 3Mieap-knockdown
SA202968A549 Non-infection 1Non-infection
SA202969A549 Non-infection 3Non-infection
SA202970A549 Non-infection 2Non-infection
SA202971A549 Non-infection 4Non-infection
Showing results 1 to 20 of 20

Collection:

Collection ID:CO002192
Collection Summary:Cells were collected using trypsin-EDTA. The cells were snap-frozen in liquid nitrogen after cell count, and subsequently stored at -80°C until lipidomic analysis.
Sample Type:Cultured cells
Volumeoramount Collected:10,000,000 cells/tube
Storage Conditions:-80℃

Treatment:

Treatment ID:TR002211
Treatment Summary:Infection of the A549 cell line was carried out by adding viral solution (Ad-Mieap) to A549 cell monolayers, and incubating at 37°C for 120 min with brief agitation every 20 min. This was followed by the addition of culture medium and the return of the infected cells to the 37°C incubator. We established a Mieap-KD cell line using LS174T. Mieap expression was inhibited in the cell line by retroviral expression of short-hairpin RNA (shRNA) against the Mieap sequence. We also established LS174T-cont cells using the retroviral vector with target sequence for EGFP. The LS174T-cont and Mieap-KD cells were incubated under normal condition.
Treatment Doseduration:A549 cells: 24 h; LS174T-cont and Mieap-KD cells: none (incubated under normal condition)
Treatment Vehicle:A549 cells: viral solution (Ad-Mieap); LS174T-cont and Mieap-KD cells: none (incubated under normal condition)
Cell Storage:stored at -80°C

Sample Preparation:

Sampleprep ID:SP002205
Sampleprep Summary:Total lipids were extracted from samples using the Bligh-Dyer method. An aliquot of the organic phase was added to an equal volume of methanol before being loaded onto a DEAE-cellulose column (Wako Chemical) pre-equilibrated with chloroform. After successive washes with chloroform/methanol (1:1, v/v), acidic phospholipids were eluted with chloroform/methanol/HCl/water (12:12:1:1, v/v), followed by evaporation to dryness to yield a residue was soluble in methanol.
Extraction Method:the Bligh-Dyer method

Combined analysis:

Analysis ID AN003461
Analysis type MS
Chromatography type Reversed phase
Chromatography system UltiMate 3000 (Thermo Fisher Scientific)
Column Waters X-Bridge C18 (150 x 1.0mm,3.5um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Plus Orbitrap
Ion Mode NEGATIVE
Units pmol/10E6 cells

Chromatography:

Chromatography ID:CH002556
Instrument Name:UltiMate 3000 (Thermo Fisher Scientific)
Column Name:Waters X-Bridge C18 (150 x 1.0mm,3.5um)
Chromatography Type:Reversed phase

MS:

MS ID:MS003223
Analysis ID:AN003461
Instrument Name:Thermo Q Exactive Plus Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:pmol/10,000,000 cells
Ion Mode:NEGATIVE
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