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MB Sample ID: SA201215
| Local Sample ID: | A549 Non-infection 4 |
| Subject ID: | SU002181 |
| 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 |
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Subject:
| Subject ID: | SU002181 |
| 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:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| A549 Non-infection 4 | SA201215 | FL024474 | Non-infection | Genotype/Treatment |
Collection:
| Collection ID: | CO002174 |
| 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: | TR002193 |
| 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: | SP002187 |
| 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 | AN003425 | AN003426 | AN003427 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | Reversed phase |
| Chromatography system | UltiMate 3000 (Thermo Fisher Scientific) | Agilent 1290 Infinity | Agilent 1290 Infinity |
| Column | Waters X-Bridge C18 (150 x 1.0mm,3.5um) | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Triple quadrupole | Triple quadrupole |
| MS instrument name | Thermo Q Exactive Plus Orbitrap | Agilent 6495 QQQ | Agilent 6495 QQQ |
| Ion Mode | NEGATIVE | POSITIVE | NEGATIVE |
| Units | pmol/10,000,000 cells | counts | counts |
Chromatography:
| Chromatography ID: | CH002533 |
| 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: | MS003188 |
| Analysis ID: | AN003425 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | pmol/10,000,000 cells |
| Ion Mode: | NEGATIVE |
