Figure 6.

Knockdown VDR induces Wnt4/β-catenin and mTOR signaling and induces proliferation of UtSMC cells. A, VDR was targeted for knockdown in UtSMC cells by infecting with lentiviruses expressing VDR-specific shRNA or scrambled control. Cell lysates were analyzed by Western blotting using anti-VDR, anti-β-catenin, anti-Wnt4, anti-p-mTOR, and anti-mTOR antibodies. β-Actin was used as loading control. B, Quantification of protein expression of above proteins were normalized to β-actin and shown. *, P < .05 as compared with control. C, Cell lysates were also analyzed by Western blotting using antifibronectin, anticollagen type 1, and anti-PCNA antibodies. D, Normalized protein levels are shown. *, P < .05 as compared with control. E, Both scrambled control and VDR knockdown cells were seeded into 12-well plates and cultured in phenol-free DMEM/F12 medium containing 10% charcoal stripped fetal bovine serum. Cultures were replenished every other day with fresh conditioned media. Cell proliferation MTT assay was performed at days 2, 3, 4, and 5. Each data point is the mean SD of triplicate wells (n = 3). *, P < .05 as compared with the corresponding control.