Figure 7. The TRIM24 bromodomain and AR-interacting LxxLL motif are required for proliferation of CRPC cells.

(A) Proliferation curves of abl cells stably expressing doxycycline (dox)-inducible shRNA targeting TRIM24 or a non-targeting (NT) control are shown. The shTRIM24 cell line was complemented with TRIM24 cDNA-rescue constructs resistant to shTRIM24, including TRIM24-WT (WT), TRIM24-L763A, L764A (L763A, L764A) and TRIM24-F979A, N980A (F979A, N980A). Cultures were treated with dox at day 0 and were grown and counted at 0, 3 and 6 days and values were plotted. Statistical analysis was performed using a two-tailed student's t-test assuming unequal variance ***p<0.005. (B) TRIM24 protein levels of the cell lines complemented with the indicated constructs were measured after 6 days of dox-treatment. (C, D) AR-specific ChIP followed by qPCR was performed at the AURKB enhancer, which is co-occupied by AR and TRIM24 (n=2). The different cell lines assayed are described in (A) and include (C) abl cells expressing shTRIM24 or shNT, and (D) abl shTRIM24 cell lines complemented with TRIM24-WT, TRIM24-L763A, L764A and TRIM24-F979A, N980A. All cell lines were treated with dox for 6 days to induce shRNA expression. The relative enrichment of the AR ChIP compared to the total input amount was set at 100% for the respective control condition. Statistical analysis was performed using a one-tailed student's t-test assuming equal variance * p<0.05, *** p<0.005. Data are represented as mean +/- SEM.

(See also Figure S5).