RARg bookmarking functions in prostate cancer (CUT&RUN)
To define the RARg complex in PCa cell models, including isogenic variants of 22Rv1, how this is targeted by miR-96, and regulates the AR cistrome, and how this is specifically enriched in G2/M cells and how these actions collectively shape the transcriptional programs associated with PCa outcomes. Overall design: CUT&RUN was undertaken with antibodies to GFP (for RARgamma), AR, H3K27ac, H3S10P and IgG in 22Rv1 isogenic cell variants.  Specifically 22Rv1 cells stably transfected with an empty vector (22Rv1-mock) or a GFP-tagged RARgamma (22Rv1-RARgamma). 22Rv1-RARgamma-TACC1 cells were generated by transient transfection of TACC1 into 22Rv1-RARgamma cells; TACC1 protein expression was sustained for ~72h. Briefly, 0.5x106 cells were treated for DHT (10nM, 6h) or CD437 (400 nM, 6h).  To synchron