Fig. 5.

CDK8/19 kinase activity is required for BMP4-induced EMT. a Effect of Senexin B on BMP4 (30 min treatment) induced SMAD1 phosphorylation at Ser206 in indicated cell lines in the absence or presence of Senexin B as indicated. Quantitation normalized to total SMAD1 for pSer206 from two independent biological trials for each cell line using LI-COR Biosciences linear range quantitation tool are presented (right graph). b Real-time quantitative PCR showing fold change in mRNA levels of SNAI1, SNAI2 transcription factors in indicated cells pre-treated with Senexin B for 30 min, then treated with BMP4 (10 nM) for 24 h. Graphs are representative of two independent biological trials done in triplicate. Error bars represent the standard error of the mean. c OvCa429 and Panc1 cells were pre-treated with Senexin B for 30 mins and plated in transwell chambers coated with Matrigel in the absence or presence of BMP4 (10 nM). Cells were allowed to invade for 18 h. (Graph) Graphical representation (right graph) of invading cells from four independent fields representing three independent biological trials. Error bars represent the standard error of the mean. d Immunofluorescence images of OvCa429, Panc1, and Py2T cells pre-treated for 30 mins with Senexin B (5 μM), then treated with BMP4 (10 nM) for 1 h followed by immunostaining with YAP1 Scale bars = 20μm. Quantitation of nuclear to cytoplasmic fluorescence intensity ratio is presented for each cell line (right graphs) *p < 0.05, **p < 0.01, ***p < 0.001