Figure 6.

Rmrp regulated the proliferation and fibrosis of MC via miR-1a-3p/JunD signaling. (A, B) The relative expression of JunD was detected after transfection with miR-1a-3p mimics and inhibitor by qRT-PCR and western blot. (C) The schematic model revealed the JunD 3’ -UTR wide type (WT) and mutant (Mut) luciferase reporter vectors. (D) The luciferase reporter assays showed that miR-1a-3p directly inhibited the luciferase activity of JunD 3’ -UTR WT, but lost efficacy when the Rmrp was up-regulated. (E, F) IHC analysis and western blot assessed JunD expression in the renal tissues of DN and normal mice (Scale bar, 50 μm). (G) The relative expression of JunD in MC after transfection with Rmrp expression vector or/and miR-1a-3p mimics by qRT-PCR. (H) The cells proliferating were performed to detect the competition effects of Rmrp and JunD siRNA by EdU (Scale bar, 50 μm). (I) The protein expression of JunD, Cyclin D1, CKD4, Col-IV and FN was analyzed after regulation the level of Rmrp expression vector, JunD siRNA or both of them by western blot. (J) The schematic diagram illustrates the mechanism of Rmrp mediated by Sp1 to mesangial cell proliferation and fibrosis through Rmrp/miR-1a-3p/JunD axis in DN. Data were represented as the mean ± SD of three independent experiments; *p < 0.05, **p < 0.01, and ***p < 0.001. ns, no significant.