Additional file 1 of (Pro)renin receptor mediates tubular epithelial cell pyroptosis in diabetic kidney disease via DPP4-JNK pathway

Additional file 1: Figure S1. Schematic diagram showing the injection sites of adeno-associated virus (AAV9-shPRR or negative control (AAV9-shNC). Figure S2. Pathological changes of glomerulus in DKD patients. A, B Representative transmission electron microscopy images of glomerular basement membrane in renal sections from DKD patients and health controls (A) and quantitative analysis of glomerular basement membrane (GBM) (B). n=5. Bar=1 μm C–F Representative immunohistochemistry staining images of WT-1 and CD31 in renal sections from DKD patients and health controls (C, E) and quantitative analysis (D, F). n=5. Bar=10μm. Data are presented as mean ± SEM of biologically independent samples. ∗∗P < 0.01. P values were determined by Student’s t-test for comparison between two groups. Figure S3. Knockdown of PRR or DPP4 alleviated high glucose induced pyroptotic morphological changes of HK-2 cells. Representative scanning electron microscope (SEM) images of HK-2 cells under different treatments. Scale bars: 20 μm. Figure S4. PRR exerted pyroptotic effects independent of Ang II in HK-2 cells. A and B Representative western blot analyses (A) and quantitative data (B) showed that blocking Ang II receptor with losartan had no significant effect on PRR overexpression induced protein expression of NLRP3, cleaved-Caspase1, GSDMD-N, IL-1β and IL-18 in HK-2 cells (n = 3). Data are presented as mean ± SEM of biologically independent samples. ∗P < 0.05, ∗∗P < 0.01, ns P>0.05. One-way ANOVA was used to analyze the data among multiple groups, followed by Tukey’s post hoc test. Figure S5. PRR promoted EMT and EndMT in the kidney of db/db mice. A and B Representative western blot analyses (A) and quantitative data (B) showed that AAV9-shPRR reduced the expression of α-SMA and Vimentin and restored the abundance of E-Cadherin and CD 31in the kidney of db/db mice (n = 6). Data are presented as mean ± SEM of biologically independent samples. ∗∗P < 0.01. One-way ANOVA was used to analyze the data among multiple groups, followed by Tukey’s post hoc test.