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. 2020 Jul 8;11:436. doi: 10.3389/fendo.2020.00436

Figure 4.

Figure 4

Smad2 and Smad3 exert opposite effects on the expression of key genes involved in enEVT differentiation. HTR-8/SVneo cells were transfected with a siRNA targeting Smad2 (siSmad2) or Smad3 (siSmad3) or a non-targeting control (NC). Total RNA or protein was extracted. (A) Validation of Smad2 siRNA (siSmad2) by qPCR. (B) Validation of siSmad3 using qPCR. (C) Validation of siSmad2 and siSmad3 by Western blotting using an antibody detecting both total Smad2 and total Smad3. (D) Knockdown of Smad2 significantly increased the mRNA levels of MMP1, IL1b, IL8, ECSCR, and VE-cad. (E) Knockdown of Smad3 significantly reduced the mRNA levels of MMP1, IL1b, IL8, ECSCR, and VE-cad. Data represent mean ± SEM (n = 3). *p < 0.05; **p < 0.01; ***p < 0.001; and ****p < 0.0001.