Fig. 3

Translation, not transcription, is responsible for AR protein level changes after R1881 treatment. A Relative change of AR mRNA levels after 6 h treatment with DMSO (CTRL), 1 µM 1 nM R1881, 1 nM R1881 + 10 µM Enzalutamide, and 1 nM R1881 + 20 µg/ml Cycloheximide in LNCaP, C4-2 and 22Rv1 cells. Relative expression levels after treatment were shown as mean ± SEM of five independent experiments. B Representative western blots of the androgen receptor (AR), AR splice variant 7 (V7), and total protein stain (TPS) after treatment with DMSO (CTRL), 1 nM R1881, 1 nM R1881 + 10 µM Enzalutamide, and 1 nM R1881 + 20 µg/ml Cycloheximide in LNCaP, C4-2 and 22Rv1 cells. Uncropped western blots are displayed in Supplementary Fig. 4A. C-E Densitometry of AR protein levels relative to TPS in LNCaP (C), C4-2 (D), and 22Rv1 (E) cells. Relative expression levels compared to CTRL after treatment were shown as mean ± SD of six independent experiments. All differences highlighted by asterisks were statistically significant (*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001)