Figure 2 | Cell Death & Disease

Figure 2

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

Figure 2

miR-558 activates the promoter activity and transcription of HPSE in an AGO1-dependent manner. (a) Scheme and sequence of the wild type (WT) and mutant (Mut) miR-558 binding site within the luciferase reporter of HPSE promoter. (b) and (c) Dual-luciferase assay showing the activity of HPSE promoter and its mutant in SGC-7901 and AGS cells transfected with empty vector (mock), miR-558 precursor, negative control inhibitor (anti-NC, 100 nmol/l) or anti-miR-558 inhibitor (100 nmol/l). (d) and (e) Western blot and real-time quantitative RT-PCR assays indicating the expression of AGO1, AGO2, AGO3, AGO4 and HPSE in gastric cancer cells transfected with mock or miR-558 precursor, and those co-transfected with scramble siRNA (si-Scb) or siRNAs specific for AGO1, AGO2, AGO3 or AGO4. (f) Dual-luciferase assay showing the HPSE promoter activity in gastric cancer cells stably transfected with mock or miR-558 precursor, and those co-transfected with si-Scb or si-AGO1. (g) ChIP and qPCR assay indicating the binding of AGO1 to HPSE promoter in gastric cancer cells treated with RNase H or RNase A. (h) ChIP and qPCR assay showing the enrichment of AGO1, H3K9me2, H3K27me3, H3K4me3 and Smad4 on HPSE promoter in SGC-7901 and AGS cells transfected with mock or miR-558 precursor, and those co-transfected with si-Scb or si-AGO1. *P<0.01 versus mock, anti-NC, mock+si-Scb or IgG

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