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. 2016 Sep 7;7(43):70404–70419. doi: 10.18632/oncotarget.11879

Figure 3. Androgen differentially regulated MYC expression.

Figure 3

A. Western blot for AR and MYC in prostatic cells. Androgen (DHT, 10 nM) treatment resulted in up-regulation of MYC in LNCaP cells, but down-regulation of MYC in NHPrE1/AR and PC3/AR cells. Lower panel is the quantification of MYC Western blot. B. Western blot for AR and MYC. NHPrE1/AR cells were cultured in androgen-depleted medium for 2 days with or without the addition of 10 nM DHT and/or 10 μM Bicalutamide (Bic). Bicalutamide treatment reversed androgen-mediated reduction of MYC. C. Analysis of MYC protein stability. Cycloheximide chase analyses were conducted using NHPrE1/AR cells to determine whether androgen treatment affected the turnover of MYC. NHPrE1/AR cells were treated with 50 μg/ml cycloheximide to block protein synthesis in the presence or absence of 10 nM DHT and harvested at different time points post treatment. Androgen treatment did not alter the stability of MYC protein in NHPrE1/AR cells. Lower panel is the semi-logarithm plot of MYC levels at different times of cycloheximide treatment. D. qRT-PCR to assess the levels of MYC mRNA in NHPrE1/EV, NHPrE1/AR, and PC3/AR cells. The expression of GAPDH was used to normalize the qPCRs. DHT treatment significantly decreased the level of MYC mRNA in NHPrE1/AR and PC3/AR cells. *p<0.05, **p<0.01, t-test.