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. 2018 Apr 16;9:312. doi: 10.3389/fphys.2018.00312

Figure 8.

Figure 8

Effects of treatments with antiandrogens on expression levels of ubiquitin E3 ligases, FBW7, MDM2, and MDM4 transcripts and effects of the siRNA-mediated inhibition of ubiquitin E3 ligases on expression levels of KCa1.1 proteins in MDA-MB-453 cells. (A–C) Real-time PCR assay for FBW7 (A), MDM2 (B), and MDM4 (C) in vehicle-, 1 μM BCT-, and 1 μM EZT-treated MDA-MB-453 cells (n = 4 for each). (D) Protein lysates of FBW7, MDM2, and MDM4 siRNA-transfected MDA-MB-453 cells (for 72 h) were probed by immunoblotting with anti-KCa1.1 (upper panel) and anti-ACTB (lower panel) antibodies on the same filter. (E) Summarized results were obtained as the optical density of KCa1.1 and ACTB band signals. After compensation for the optical density of the KCa1.1 protein band signal with that of the ACTB signal, the KCa1.1 signal in the vehicle control was expressed as 1.0 (n = 4 for each). Expression levels were expressed as a ratio to ACTB. Results are expressed as means ± SEM. **p <0.01 vs. the vehicle control or control siRNA (si-cont).