Fig. 2
c-Met and TGFβ signalling in HGF-mediated EMT of NBT-II carcinoma cells. a Bar graph displays ratio of phosphorylated proteins at 2 h post HGF compared with control. b Western blot analysis of NBT-II cells treated with escalating doses of HGF at various concentrations for 2 h. Lysates are probed with indicated antibodies. c Western blot analysis of NBT-II cells treated with 5 ng/ml of HGF. Lysates were collected at denoted time points and probed with indicated antibodies. d Western blot analysis of NBT-II cells treated with 5 ng/ml of HGF in the absence or presence of c-MET inhibitor JNJ38877605. Lysates were collected at denoted time points and probed with indicated antibodies. e Western blot analysis of nuclear (n) and cytoplasmic (c) fractions isolated from NBT-II cells treated with HGF for 2 h with or without TβRI inhibitor (A83-01, 8 μM). Lysates were collected at denoted time points and probed with indicated antibodies. f Bar chart displays ratio of pSMAD2 band intensity in nuclear (n) fractions compared with total SMAD2 (nuclear + cytoplasmic) fractions isolated from NBT-II cells treated with HGF for 2 h with or without TβRI inhibitor (A83-01, 8 μM). Bars represent mean ± SD of three independent experiments. A two-tailed Student’s t test compares the treated cell populations, **P < 0.01, ***P < 0.001. g Immunofluorescence images of SMAD2-stained cells transfected with either control vector or TβRI-dominant negative (DN) mutant construct with or without HGF. Scale bars: 10 μm. h Scatter plot of 245 pathways combined enrichment score (x axis) and p value in −log10 scale (y axis) from Enrichr. Pathways related to TGFβ are denoted in red. Dotted line indicates p = 0.05. The adjusted p value is the Benjamini–Hochberg corrected p value from hypergeometric test
