Fig. 2: CRPC models display increased durability of KLF5 induction by androgens.

a KLF5 mRNA measured by RT-PCR in LNCaP cells cultured in androgen-deplete medium supplemented with 1 nM DHT for indicated time-points. n = 6, mean ± 95% CI from 2 biological replicates in technical triplicate. b KLF5 protein measured by western blot using the same conditions as a. ERK2 is a loading control. c KLF5 mRNA measured by RT-PCR in VCaP cells as in a. n = 6, mean ± 95% CI from 2 biological replicates in technical triplicate. d KLF5 protein measured by western blot using the same conditions as in c. ERK2 is a loading control. e KLF5 protein measured by western blot in VCaP cells treated with 1 nM mibolerone (Mib) as in d. One additional replicate experiment was performed that yielded a comparable result. f KLF5 protein measured by western blot in C4-2B cells treated as in b. One additional replicate experiment was performed that yielded a comparable result. g KLF5 mRNA measured by RT-PCR in LNCaP and LNCaP-derived CRPC sub-lines 16D, 49F, and 42D cells treated as in a. n = 6, mean ± 95% CI from three biological replicates in technical duplicate. h Schematic of LNCaP cell culture conditions for i. enz = enzalutamide, DMSO = vehicle control. i KLF5 mRNA measured by RT-PCR in cells cultured as in i. n = 6, mean ± 95% CI from 2 biological replicates in technical triplicate. j KLF5 protein measured in R1-AD1 cells by western blot using the same conditions as a. One additional replicate experiment was performed that yielded a comparable result. k KLF5 and AR proteins measured by western blot using LNCaP cells cultured in androgen-deplete medium (CSS), LNCaP cells stimulated 8 h with 1 nM DHT as in a, or indicated cell lines grown in their respective standard medium conditions. Actin is loading control. One additional replicate experiment was performed that yielded a comparable result.