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. 2019 May 17;11(5):691. doi: 10.3390/cancers11050691

Figure 1.

Figure 1

Effect of RAC1B knockdown (KD) and knockout (KO) on activin receptor-like kinase 5 (ALK5) expression in Panc1 cells. (A) Panc1-RAC1B-KD cells were generated by transfecting Panc1 cells twice with 50 nM of either control (Co) siRNA or RAC1B siRNA. Forty-eight hours after the second transfection, the cells were treated with transforming growth factor (TGF)-β1 for 24 h and subsequently processed for qPCR of ALK5 and TATA-box-binding protein (TBP) (left-hand panel) or immunoblot analysis of ALK5, RAC1B, and heat shock protein (HSP)90 (right-hand panel). The graphs show quantification of ALK5 data normalized to those for TBP (ΔΔCt values from qPCR) or HSP90 (densitometric values from immunoblots) and represent the mean ±SD from three independent experiments. (B) Panc1-RAC1B-KO and vector control cells were treated with TGF-β1 for 24 h and processed for either qPCR of ALK5 and β-actin (left-hand panel) or immunoblot analysis of ALK5 and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (right-hand panel). Expression of RAC1B in these cells is shown in Figure S1. The graphs indicate the normalized data (mean ±SD, n = 3). The asterisks indicate significance (student’s t-test). V, vector control; ns, not significant.