Fig. 4: Nestin induced the mitophagy increased in lupus nephritis, and regulated the phosphorylation of nephrin by mitophagy.

a IF assay showed that the expression of nestin, LC3, p62 and PINK1 in MPCs treated with LN plasma. Scale bars: 10 μm. b, c Western blot assay showed the expression of Atg5, Atg12, Atg16, PINK1, LC3 and p62 in the MPCs exposed to LN plasma. *P < 0.05 vs. 0 h group (n = 3). d–f Western blot assay showed the expression of Atg5, Atg12, Atg16, PINK1, LC3, p62, nestin, nephrin and p-nephrin (Y1217) in the MPCs, which were pretreated with 3-MA to inhibit the autophagy and exposed to LN plasma for 24 h. *P < 0.05 vs. control plasma group, ^#P < 0.05 vs. LN plasma group, n.s. no significance (n = 3). g, h Knockdown of the expression of PINK1 effectively by siPINK1 in MPCs exposed to LN plasma for 24 h, Western blot assay showed the expression of PINK1, nestin, nephrin and p-nephrin (Y1217). *P < 0.05 vs. control plasma group, ^#P < 0.05 vs. LN plasma + siNC group, n.s. no significance (n = 3). i, j Western blot assay showed the downregulation of nestin affected the expression of Atg5, Atg12, Atg16, PINK1, LC3 and p62 in the MPCs exposed to LN plasma for 24 h. *P < 0.05 vs. control plasma group, ^#P < 0.05 vs. LN plasma + shNC group (n = 3). k, l Western blot assay showed the downregulation of nestin affected the expression of Atg5, Atg12, Atg16, PINK1, LC3 and p62 in the MPCs. *P < 0.05 vs. control FBS group (n = 3). m, n Western blot assay showed the overexpression of nestin affected the expression of Atg5, Atg12, Atg16, PINK1, LC3 and p62 in the MPCs exposed to LN plasma for 24 h. *P < 0.05 vs. control plasma group, ^#P < 0.05 vs. LN plasma + NC group (n = 3). o, p Western blot assay showed the overexpression of nestin affected the expression of Atg5, Atg12, Atg16, PINK1, LC3 and p62 in the MPCs. *P < 0.05 vs. control FBS group (n = 3). Bonferroni’s correction was performed to analyze statistical significance. Values are mean ± SEM.