Fig. 4

Enzalutamide induces GATA2-dependent CDC6 upregulation and further promotes EMT upon acquiring therapy resistance. A Western blotting and densitometry in C4-2B and PC-3 cells transfected with the indicated siRNAs and/or treated with DMSO or enzalutamide. Cell lysates were probed with the indicated antibodies. CDC6 is stabilized either upon GATA2 depletion or enzalutamide treatment. B qPCR for GATA2 mRNA levels validating successful GATA2 KD in A. Endogenous GATA2 levels are downregulated in response to therapy in already enzalutamide-resistant cells. See also Fig. S5. C qPCR for EMT markers CDH1, ZEB1 and SNAI1 in the indicated conditions of cells from A. GATA2 depletion or enzalutamide treatment induce EMT marker expression. Enzalutamide was used at a 10 μM concentration. **P < 0.01 and ***P < 0.001, of Student’s t-test. Error bars indicate s.e.m. Data shown are representative of at least 3 biological experiments (n ≥ 3)