SPOP mass spec in different treatment

ABSTRACT: 293T cells were transfected with S, Flag, and Biotin-binding-protein-(streptavidin)-binding-peptide (SFB) triple-tagged backbone vector or SFB-tagged SPOP. Twenty-four h after transfection, control cells were treated with DMSO (Group 1) and SFB-SPOP transfected cells were treated with DMSO (Group 2) or DNA damaging agents including mitomycin C (MMC, 1 µM) (Group 3), camptothecin (CPT, 50 nM) (Group 4), cisplatin (10 µM) (Group 5) and etoposide (10 µM) (Group 6) for 24 h. The cells were lysed by NETN buffer (20 mM Tris-HCl, pH8.0, 100 mM NaCl, 1mM EDTA, 0.5% Nonidet P-40) with 50 mM β-glycerophosphate, 10 mM NaF, and 1μg/mL pepstatin-A at 4°C for 3 h, followed by tandem affinity purification using streptavidin beads and S tag beads and LC-MS. A total of six groups of cell lysate were analyzed (n=6), including one group of cells transfected with empty vector and treated with DMSO as a control.

ORGANISM(S): Homo Sapiens

SUBMITTER: Jian Ma  

PROVIDER: PXD028210 | iProX | Mon Sep 06 00:00:00 BST 2021

REPOSITORIES: iProX