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

FIGURE 2.

FIGURE 2

Gremlin via VEGFR2 induces EMT in tubular epithelial cells. (A) Cells (HK2 cell line) were preincubated with the VEGFR2 kinase inhibitor SU5416 (5 μM) or vehicle before stimulation with Gremlin (10 ng/ml) for 72 h. By confocal microscopy changes in epithelial phenotype and EMT-markers were evaluated. Vimentin is shown in green and Cytokeratin in red. Figure shows a representative experiment of two done by confocal microscopy. (B) Changes in EMT-related markers were quantified by western blot Gremlin diminished the levels of the epithelial markers E-Cadherin and Cytokeratin and increased the expression of mesenchymal markers, α-SMA and vimentin. All these changes were prevented by VEGFR2 inhibition. (C) Gene expression was evaluated 24 h after Gremlin stimulation. Total cell RNA was isolated to assess mRNA levels of VIMENTIN, and E-CADHERIN by quantitative real-time PCR. Figure shows mean ± SEM of six independent experiments. ∗p < 0.05 vs. control; #p < 0.05 vs. Gremlin.