Proteomics

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CDKN1B (p27kip1) enhances drug tolerant persister CTCs by restricting polyploidy following mitotic inhibitors


ABSTRACT: BRx82 and BRx142 were cell cultures derived from breast cancer circulating tumor cells and propagated ex-vivo. To study early changes that transpire under mitosis inhibiting chemotherapy, these cultures were subjected to a 16 hours exposure to docetaxel (10nM or DMSO as vehicle control) and then cultured for a total of 4 days. The experiments were done using an Orbitrap Fusion (proteomics) or an Orbitrap Fusion Lumos (phosphoproteomics) mass spectrometer. Multiplexing was achieved using either TMT10 reagents and the SPS-MS3 method. Basic pH reversed-phase chromatography (bRPLC) was used for off-line pre-fractionation; 12 fractions were analyzed for proteome mappings (PMID: 26700037) and 24 fractions sample for phosphoproteome mappings (PMID: 31606085). Phosphoproteome mappings were done using both HCD fragmentation with Orbitrap fragment ion detection and CID fragmentation with ion trap fragment ion detection (PMID: 29487189, PMID: 31606085). RAW files from the proteome mapping experiment are named HorwitzMaheswaranHaber_proteome_fraction01 to 12. RAW files from the phosphoproteome mapping experiment are named TMT labeling was the same for the proteomics and the phosphoproteomics experiments: BRx82_DTX-_#1 (129c) BRx82_DTX-_#2 (130n) BRx82_DTX+_#1 (127c) BRx82_DTX+_#2 (128n) BRx142_DTX-_#1 (126) BRx142_DTX-_#2 (127n) BRx142_DTX+_#1 (128c) BRx142_DTX+_#2 (129n)

INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Fusion

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Wilhelm Haas   Shyamala Maheswaran   Daniel Haber  

PROVIDER: MSV000094243 | MassIVE | Tue Mar 05 20:31:00 GMT 2024

REPOSITORIES: MassIVE

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