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. 2023 Nov 18;20(5):1032–1053. doi: 10.1080/15548627.2023.2281156

Figure 5.

Figure 5.

Autophagy deficiency inhibits MAPK1/ERK2-MAPK3/ERK1 activation in maladaptive renal tubules and this inhibition correlates with SQSTM1/p62 aggregation and sequestration of MAPK1/ERK2-MAPK3/ERK1 in iRT-atg7 KO mice. WT and iRT-atg7 KO mice underwent sham operation or 30-min unilateral renal ischemia followed by reperfusion for up to 4 weeks. Left kidneys were harvested. (A) immunoblot of p-MAPK1/ERK2-MAPK3/ERK1 (Thr202/Tyr204), MAPK1/ERK2-MAPK3/ERK1, LC3B, SQSTM1/p62, p-ELK1 (Ser383) and ELK1. (B) densitometry of p-MAPK1/ERK2-MAPK3/ERK1 (Thr202/Tyr204) and SQSTM1/p62 immunoblot (sham: n = 4; UI30R 2 w: WT n = 7, KO n = 7; UI30R 4 w: WT n = 6, KO n = 7). (C) correlation analysis of protein fold changes between p-MAPK1/ERK2-MAPK3/ERK1 (Thr202/Tyr204) and SQSTM1/p62. (D) co-immunofluorescence of MAPK1/ERK2-MAPK3/ERK1 (red) and SQSTM1/p62 (blue). Scale bar: 20 µm. (E) quantification of SQSTM1/p62 aggregates with or without MAPK1/ERK2-MAPK3/ERK1 co-staining in post-ischemic iRT-atg7 KO mice (n = 6). Data in (B) and (E) presented as mean ± SEM. For statistics, two-way ANOVA with multiple comparisons was used for (B). Pearson correlation analysis followed by simple linear regression was used for (C). 2-tailed, unpaired Student t-test and fraction of total analysis were used for (E).