Figure 3 | Cell Death & Disease

Figure 3

From: miRNA-558 promotes gastric cancer progression through attenuating Smad4-mediated repression of heparanase expression

Figure 3

miR-558 attenuates the Smad4-mediated repression of HPSE transcription in gastric cancer cells. (a) Western blot showing the expression of Smad4 and HPSE in primary stomach epithelial HPSEC cells, SV40-immortalized normal gastric epithelial GES-1 cells and gastric cancer cell lines (MKN-45, SGC-7901, AGS and MKN-28). (b) and (c) Western blot and real-time quantitative RT-PCR assays indicating the protein and transcript levels of Smad4 and HPSE in MKN-45 and SGC-7901 cells stably transfected with empty (mock) or Smad4. (d) and (e) Western blot and real-time quantitative RT-PCR assays showing the protein and transcript levels of Smad4 and HPSE in SGC-7901 and AGS cells stably transfected with scramble shRNA (sh-Scb) or shRNA specific for Smad4 (sh-Smad4). (f) and (g) Dual-luciferase assay indicating the HPSE promoter activity in MKN-45 and SGC-7901 cells stably transfected with empty vector (mock) or Smad4, and those co-transfected with miR-558 precursor or siRNA specific for AGO1 (si-AGO1). (h) and (i) ChIP and qPCR assay showing the binding of Smad4 to the HPSE promoter in gastric cancer cells, and those stably transfected with empty vector (mock), Smad4 or miR-558 precursor. *P<0.01 versus mock, sh-Scb, mock+ si-Scb or IgG

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