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. 2017 Aug 18;18(8):1795. doi: 10.3390/ijms18081795

Figure 3.

Figure 3

Mitogen-activated protein kinases (MAPKs) are signaling intermediates required for Rho-kinase-mediated Jag1 induction. (A) Differentiated E11 podocytes were pretreated with Smad3 inhibitor (SIS3, 10 µM) and then stimulated with TGF-β (5 ng/mL) for 8 h. Jag1 mRNA was analyzed by real-time quantitative PCR; (B) Podocytes were pretreated with Y-27632 (10 µM) before stimulation with TGF-β (5 ng/mL) for 30 min. Cell lysates were subjected to Western blotting. A representative blot of three independent experiments is shown; (C) Podocytes were stimulated with TGF-β for 8 h with or without pretreatment of MAPK inhibitors (50 µM). RNA was extracted, and Jag1 mRNA was analyzed by real-time quantitative PCR, with GAPDH mRNA as the internal standard. * p < 0.05 vs. DMSO with TGF-β; (D) Podocytes were pretreated with Y-27632 (10 µM) and then stimulated with TGF-β (5 ng/mL) for 30 min. Equal amounts of cell lysate were subjected to Western blotting using MAPKs antibodies. A representative blot of three independent experiments is shown. The data are presented as means ± SD.