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Quantitative proteomic profiling of HCC1954 cells treated with a CDK12 degrader

ABSTRACT: HCC1954 cells growing at 80% confluency in a 10-cm dish were treated with (R)-2-(4-(9-ethyl-6-(((6-(furan-3-yl)pyridin-3-yl)methyl)amino)-9H-purin-2-yl)morpholin-3-yl)ethan-1-ol (a CDK12 degrader) at 300 nM (0.5% in DMSO) or DMSO as a control for 2 hours. Each was carried out in triplicates. The cells were disrupted in a lysis buffer containing 2% SDS, 2 mM MgCl2, 50 mM triethylammonium bicarbonate, 1x HALT phosphatase and protease inhibitors (ThermoFisher Scientific, Waltham, MA) using Bioruptor Sonicator (Diagenode, Denville, NJ). Proteins were recovered and proteolyzed with trypsin on S-trap columns (PROTIFI, Fairport, NY). The resultant peptides were labeled with 6-plex TMT isobaric reagents (triplicates for 2 conditions), mixed altogether, subfractionated by basic reversed phase high performance liquid chromatography, and analyzed by LC-MS/MS (ThermoFisher, Q-Exactive). The LC-MS/MS data were processed by using Proteome Discoverer v2.4.

INSTRUMENT(S): Q Exactive

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

SUBMITTER: Cheolju lee

PROVIDER: MSV000092579 | MassIVE | Wed Aug 02 23:22:00 BST 2023

SECONDARY ACCESSION(S): PXD044313

REPOSITORIES: MassIVE

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