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. 2019;10(5-6):134–149. doi: 10.18632/genesandcancer.194

Figure 3. Effect of gene knockdown on R-Smad mediated transcriptional activity.

Figure 3

A. Relative expression of phospho Smad2/3, Total Smad2/3 and TMEPAI in MDA-MB-231 cells expressing Con shRNA, Smad2 shRNA and Smad3 shRNA in the absence or presence of TGF-β (2 ng/ml), B. Relative levels of TMEPAI mRNAs in human mammary epithelial cells (HME) and MDA-MB-231 cells that are expressing Con shRNA, TMEPAI shRNA (TMKD), Smad2 shRNA (Smad2KD) or Smad3 shRNA (Smad3KD) in the absence or presence of TGF-β (2 ng/ml), C. 12XCAGA-Luc reporter activity in MDA-MB-231 cells expressing Con ShRNA, Smad2 shRNA, Smad3 shRNA both in the absence or presence of TGF-β (2 ng/ml). D. 3XARE-Luc reporter activity in MDA-MB-231 cells expressing Con ShRNA, Smad2 shRNA, Smad3 shRNA both in the absence or presence of TGF-β (2 ng/ml). E. 12xCAGA-Luc reporter activity in MDA-MB-231 cells expressing Con ShRNA (231_CON) and TMEPAI shRNA (231_TMKD) in the absence or presence of TGF-β (2 ng/ml). F. 3XARE-Luc reporter activity in MDA-MB-231 cells expressing Con ShRNA (231_CON) and TMEPAI shRNA (231_TMKD) in the absence or presence of TGF-β (2 ng/ml). G. Schematic representation of three different size TMEPAI promoters tagged with luciferase reporter (−2251-Luc, −973-Luc, −301-Luc) and corresponding luciferase reporter activities in MDA-MB-231 cells in the absence or presence of TGF-β (2 ng/ml). H. TMEPAI promoter activity in MDA-MB-231 cells expressing con shRNA, Smad2 shRNA, and Smad3 shRNA in the absence or presence of TGF-β (2 ng/ml) using 2251 bp TMEPAI promoter (−2251-Luc).