Figure 5. RXRA S427F generates PPARD-dependent urothelial growth.
(A) Retrovirally transduced organoids from Figure 4 were plated for 7 days in standard media and then treated with the indicated PPARD antagonists (1000 nM ST-247, GSK0660) or PPARG antagonist (100 nM T0070907) for 2 days. Expression of PPAR targets was determined by RT-qPCR in triplicate ±SD and comparison is by Student’s t-test to the RXRAS427F DMSO condition. (See also Figure 5—figure supplement 1)) (B-E) CellTiter-Glo growth assay of indicated organoid lines treated with indicated drugs. Plotted is mean signal ±SEM from three independent experiments, each performed using triplicate organoid wells. MCB6C is an organoid line we derived from a carcinogen-induced bladder tumor and lacks RXRA mutation. Organoids were cultured in media without EGF except for panel E where inclusion of EGF is indicated. Comparison is to DMSO condition using paired t-test.
Figure 5—figure supplement 1. Aggregated RT-qPCR from three distinct experiments as described in Figure 5A with each data point from each experiment plotted with mean indicated.
