Skip to main content
. 2020 Nov 29;12(12):3570. doi: 10.3390/cancers12123570

Figure 4.

Figure 4

Effects of siRNA-mediated inhibition of TGFβ1 or RAC1B on genes involved in EMT and cell motility. (A) Panc1 or MDA-MB-231 cells were transiently transfected twice with 50 nM of either ctrl siRNA, TGFβ1 siRNA, RAC1B siRNA, or a combination of both, as indicated. Forty-eight h later, cells were processed for qPCR analysis of CDH1 and SNAI1, and GAPDH as an internal control. (B) Panc1TGFB1KD cells were subjected to immunoblot analysis of ECAD, SNAIL, RAC1B, and HSP90 as a loading control. (C) As in (A), except that Panc1TGFB1KD cells were subjected to amplification of SMAD3 mRNA and Panc1SMAD3KD cells subjected to amplification of TGFβ1 mRNA. (D) Immunoblot analysis of SMAD3 in Panc1TGFB1KD cells. (E) Immunoblot analysis of ECAD in Panc1SMAD3KD cells. Data in (A,C) represent the mean ± SD of GOI after normalization with the housekeeping genes (TBP or GAPDH) from three independent transfection experiments. The graphs below the blots in panels (B,D,E) depict data quantification based on densitometric readings from band intensities after normalization with those for HSP90 (mean ± SD; B, D: n = 5, E: n = 4). The asterisks indicate significant differences (* p < 0.05; ** p < 0.01; *** p < 0.001).