Figure 5.

miR-27a targets GFPT2 and inactivates the TGF-β/Smad2/3 signaling pathway. A, intersected target mRNAs of miR-27a predicted on four bioinformatic systems StarBase, TargetScan, miRDB and PicTar; B, binding relationships between miR-27a and GFPT2 or NAFSC mRNA examined through dual-luciferase reporter gene assays (two-way ANOVA, ** p < 0.01 vs. GFPT2-MUT); C, binding relationship between miR-27a and GFPT2 validated through a biotin-labeled RNA pull-down assay (one-way ANOVA, ** p < 0.01 vs. Biotin-NC); D-E, mRNA (d) and protein (e) expression of GFPT2 in cells determined by RT-qPCR and western blot analysis, respectively (one-way ANOVA, ** p < 0.01 vs. control, ^## p < 0.01 vs. H/R, ^&& p < 0.01 vs. Mock, ^@@ p < 0.01 vs. ML264 + inhibitor); F, expression of TGF-β1 and phosphorylation of Smad2/3 in cells determined by western blot analysis (one-way ANOVA, ** p < 0.01 vs. control, ^## p < 0.01 vs. H/R, ^&& p < 0.01 vs. Mock, ^@@ p < 0.01 vs. ML264 + inhibitor). Data were exhibited as mean ± SD from at least three independent experiments