Journal: Cell Death & Disease
Article Title: PRDM1 restricts bladder cancer progression and enhances chemosensitivity by suppressing OTUD6A-mediated deubiquitination of CDC6
doi: 10.1038/s41419-026-08498-3
Figure Lengend Snippet: A Expression levels of ubiqutination-related protein (OTUD6A, FZR1, CDT2 and Cyclin F) in the PRDM1-knockdown T24 and UMUC3 cells were determined by Western blot. B Indicated T24 cells transfected with Flag-OTUD6A plasmids/empty vectors, Myc-CDC6 and HA-Ub were treated with MG132 for 6 h. Whole-cell lysates were prepared and subjected to IP with an anti-Myc antibody. The immunoprecipitates were analysed by Western blot. C The protein levels of PRDM1, CDC6 and OTUD6A in the indicated T24 and UMUC3 cells were determined by Western blot. D , E CCK-8 assays ( D ) and EdU incorporation assays ( E ) were used to examine the proliferation of the indicated T24 cells. F The indicated T24 tumors were weighed. G Representative IHC images indicating Ki-67, PRDM1, CDC6 and OTUD6A expression in the indicated T24 tumors are shown. H Apoptosis was measured by TUNEL assays in the indicated T24 cells treated with or without gemcitabine for 48 h. I The amount of DNA strand breaks was quantified by alkaline comet assays in the indicated T24 cells treated with or without gemcitabine for 48 h. J ATR-Chk1 pathway protein levels in the indicated T24 cells treated with or without gemcitabine for 6 h were determined by Western blot. K An image of subcutaneous tumors formed by the indicated T24 treated with gemcitabine is shown. L The indicated T24 tumors were weighed. M The protein levels of PRDM1, OTUD6A and CDC6 in the indicated UMUC3 GR cells were determined by Western blot. N Growth curves of the subcutaneous indicated UMUC3 GR tumors treated with gemcitabine are shown. O The indicated UMUC3 GR tumors were weighed. All quantitative analyses were based on three independent experiments. The error bars indicate the SDs. ** P < 0.01, *** P < 0.001, n.s. not significant, based on two-tailed Student’s t test.
Article Snippet: T24 cell line were obtained from ATCC and cultured in RPMI-1640 medium (Gibco, 11875093).
Techniques: Expressing, Knockdown, Western Blot, Transfection, CCK-8 Assay, TUNEL Assay, Two Tailed Test