Figure 3. AR positively regulates CCRK transcript and protein expression in liver and HCC cells.
(A) Ectopic AR expression increased CCRK expression. AR and CCRK expression were detected by Western blot following transient transfection. β-actin was used as a loading control. The relative CCRK transcript level was detected in LO2 immortal liver and SK-Hep1 HCC cells by quantitative RT-PCR. GAPDH was used as an internal control. (B) Silencing AR expression downregulated CCRK transcript and protein expression in Huh7 and PLC5 HCC cells. **P < 0.01; ***P < 0.001. (C) Ectopic AR expression induced perinuclear CCRK localization. Double immunofluorescence staining of AR and CCRK was performed in LO2 and SK-Hep1 cells transiently transfected with AR-expressing vector or empty vector. The nuclei were counterstained with DAPI. (D) Localization of AR and CCRK in Huh7 and PLC5 cells following RNA interference. Original magnification, ×400. Data are presented as mean + SD of 3 independent experiments.