Figure 5 TIMP3 was identified as a target gene of miR-27a-3p. (A) Potential binding sites between miR-27a-3p and TIMP3 were predicted by DianaTools-microT_CDS. (B, C) TIMP3 3′UTR WT or TIMP3 3′ UTR MUT together with miR-27a-3p, miR-NC, in-miR-27a-3p or in-miR-NC were transfected into podocytes and then the luciferase activity in podocytes was measured by dual-luciferase reporter assay. (D) The protein level of TIMP3 in HG-cultured podocytes and NC groups was measured via western blot assay. (E) After podocytes were transfected with miR-NC or miR-27a-3p and treated with HG, the level of TIMP3 protein was detected through Western blot assay. (F) Podocytes were transfected with pcDNA, 4930556M19Rik, 4930556M19Rik+miR-27a-3p or 4930556M19Rik+miR-NC and exposed to HG, then the protein level of TIMP3 was examined by western blot assay. * P<0.05.