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. 2020 Jul 28;12(8):2092. doi: 10.3390/cancers12082092

Figure 4.

Figure 4

MF-15 reduces the expression of androgen-regulated genes, diminishes PSA production and decreases the expression of AR-FL and AR-V7. (A) Expression of the androgen-regulated genes PSA and FKBP5 was determined with real-time qPCR following treatment of 22Rv1 cells with the indicated drugs over 3 days. After normalization to the housekeeping gene HMBS, fold change expression was calculated to the mock control for each individual experiment. (B) PSA levels were measured in the cell culture supernatant after treatment of 22Rv1 cells with 10 µM MF-15 on a Cobas 8000 instrument (Roche) and calculated as percentage of mock control (DMSO). (C) Expression of AR-FL, AR-V7, and AKR1C3 was assessed by real-time qPCR in 22Rv1 cells treated with MF-15 or DMSO over 3 days. Expression of each gene was normalized to HMBS and expressed as fold change to the mock control. (D) Western blot analysis of AR-FL, AR-V7 and AKR1C3 was performed as described under material and methods. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control. Representative images of blots of three individual experiments were taken with Image Studio software (Li-Cor). Data represent the mean ±SEM from three independent experiments. (statistical comparison to mock control: * p< 0.05, ** p < 0.01, *** p < 0.001; statistical comparison to R1881: # p< 0.05, ## p < 0.01, ### p < 0.001).