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. 2018 Jun 4;42(3):1484–1494. doi: 10.3892/ijmm_2018.3714

Figure 2.

Figure 2

TGF-β1 promoted the mRNA expression of NGF in SCDC2 cells in Smad2/3-dependent and p38 MAPK-dependent manners. Effects of (A) SIS3 (10 µM), and (B) SB203580 (10 µM) on expression of NGF mRNA were evaluated as described in Materials and methods. Data represent the mean ± standard deviation (n=6). *P<0.05. (C) Phosphorylation status of Smad2/3 and p38 MAPK in cells stimulated with TGF-β1 (10 ng/ml) for the indicated times, evaluated using western blot analysis. (D) After 24-h starvation, cells were pretreated with Smad3 inhibitor SIS3 (10 µM) for 30 min and then treated with or without TGF-β1 (10 ng/ml) for 30 min, and the status of nuclear translocation of Smad2/3 following TGF-β1 stimulation was examined using immunofluorescence analysis (×200 magnification; scale bar, 50 µm). (E) Phosphorylation status of MAPKAPK-2 evaluated using western blot analysis in cells stimulated with TGF-β1 (10 ng/ml) and/or with the inhibitor SB203580. (F) Effect of SP600125 (10 µM) on expression of NGF mRNA was evaluated as described in Materials and methods. Data represent the mean ± standard deviation (n=6). *P<0.05. TGF, transforming growth factor; NGF, nerve growth factor; SCDC, single cell-derived culture; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; MAPK, mitogen-activated protein kinase; MAPKAPK-2, MAPK-activated protein kinase 2.