Fig. 2

Effect of Risa overexpression and Risa inhibition on HG-induced podocyte autophagy damage. a Expression of LC3B was detected by immunofluorescence at different observation times in HG-induced MPCs. b Expression of LC3B, Beclin-1 and p62 was detected by Western blotting. The bar graph shows the quantification of LC3B II/I. c Expression of GSK3β, p-GSK3β, WT-1, NPHS2 and Desmin was detected by Western blotting. d Expression of Beclin-1 and LC3B was examined by qRT-PCR in the mouse groups and MPC groups (n = 10). e Expression of GSK3β, p-GSK3β, Beclin-1, LC3B and p62 was examined by Western blotting. The bar graph shows the quantification of LC3B II/I. f Immunofluorescence staining analysis of LC3B (green) expression, the nuclei were stained with PI (red). g Autophagosomes (white arrows) were observed by TEM. The mean numbers of autophagosomes were counted. Values are expressed as mean ± SD of three independent experiments. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 vs. the previous time group or db/m group or NG and HM groups, ^###P < 0.001 vs. db/db and db/db + Risa-vehicle groups or HG and HG + Risa-vehicle groups. MPCs mouse podocyte cells, Risa lncRNA AK044604, AAV adeno-associated virus, GAPDH glyceraldehyde-3-phosphate dehydrogenase, GSK3β glycogen synthase kinase 3β, HG high glucose, HM high mannitol, LV lentivirus, NG normal glucose