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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Mol Cancer Res. 2014 May 13;12(8):1166–1180. doi: 10.1158/1541-7786.MCR-13-0395

Figure 1. Hic-5 knockdown reduces basal and TGF-β-induced VDR mRNA and protein expression.

Figure 1

A, B. Scr and shHic-5 cells were serum-starved for 2 hr, then treated for 6 hr with 0 or 3.5 ng/mL TGF-β1. mRNA was extracted for cDNA synthesis and RT-qPCR probing for Hic-5 (A), VDR (B), and GAPDH expression. Comparisons were made using the comparative Ct (ddCt) method. TGF-β1 induced VDR transcription in both Scr and shHic-5 cells. Data were analyzed with two-way ANOVA with mixed models, followed by cellmeans post-test. Bars represent mean±SEM of six independent experiments normalized to basal expression in Scr cells. * p < 0.05, ** p < 0.01. C. Scr and shHic-5 cells were serumstarved for 2 hr, then treated for 24 hr with 0 or 10 ng/mL TGF-β1. Blot is representative of three independent experiments. D. Western blot results were analyzed by densitometry in ImageJ. TGF-β1 induced VDR expression in both Scr and shHic-5 cells. Basal expression of VDR in shHic-5 was below detectable levels, so the data was not transformed. A two-way ANOVA with mixed models followed by cell-means post-test were performed. Bars represent mean±SEM of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.