Fig. 4. Podocyte-specific METTL14 deletion promoted autophagy and inhibited apoptosis and inflammation in mice with ADR nephropathy.
Wild-type (METTL14fl/fl/Cre−) mice and podocyte-specific METTL14 knockout (METTL14fl/fl/Cre+) mice were injected with saline or ADR (19.5 mg/kg). A Representative immunoblots and quantitative analysis of autophagy-related proteins (P62 and LC3 II/LC3 I) in the renal cortex from different groups of mice (n = 6). B Representative electronic microscopy images and statistical analysis of typical autophagosomes (red arrow) in each group. Scale bar, 500 nm (n = 4). C Representative immunofluorescence images of TUNEL and WT1 double staining on kidney sections from different groups of mice. Nuclei were stained with DAPI (blue). Green fluorescence indicates TUNEL-positive nuclei. Red fluorescence represents WT1-positive nuclei. Scale bar, 20 μm. D Statistical analysis of apoptotic podocytes with TUNEL-positive and WT1-positive staining in kidney sections from each group (n = 6). E Representative immunoblots of proteins associated with apoptosis in the renal cortex of each group. F Quantitative analysis of Bcl2 and cleaved caspase-3 in these groups (n = 6). G Quantitative RT-PCR analysis of inflammatory cytokines mRNA levels (MCP-1, IL-6 and TNF-α) in groups of mice with different treatment (n = 6). *P < 0.05, **P < 0.01 vs. METTL14fl/fl/Cre− mice injected with saline, #P < 0.05, ##P < 0.01 vs. METTL14fl/fl/Cre− mice with ADR injection. Data are presented as mean ± SEM.