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. 2018 Oct 17;9:1195. doi: 10.3389/fphar.2018.01195

FIGURE 3.

FIGURE 3

VEGFR2 gene silencing blocked Gremlin-induced EMT related changes in tubular epithelial cells. Sub-confluents HK2 cells were transfected with a siRNA against VEGFR2 or siRNA scrambled during 48 h. Then, cells were stimulated or not with 10 ng/mL Gremlin for 72 h. (A) Changes in the representative marker of activated myofibroblasts α-SMA was evaluated by confocal microscopy. To see cell density the nuclear marker DAPI was used. Figure shows a representative experiment of two done by confocal microscopy. (B) Gene expression was evaluated in silenced cells 24 h after Gremlin stimulation. Total cell RNA was isolated to assess mRNA levels of VIMENTIN, and E-CADHERIN by quantitative real-time PCR. Data are expressed as mean ± SEM of five independent experiments. ∗p < 0.05 vs. control untransfected; #p < 0.05 vs. untreated control siRNA-transfected cells; $p < 0.05 vs. Gremlin-treated control siRNA-transfected cells; &p < 0.05 vs. Gremlin-treated untransfected cells.