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

Figure 7.

Figure 7.

Inhibition of SQSTM1/p62 aggregation further enhances MAPK1/ERK2-MAPK3/ERK1 phosphorylation in TGFB1-treated mouse proximal tubular cells. (A) structural domains in SQSTM1/p62 protein. (B) BUMPT cells stably expressing GFP-SQSTM1/p62 WT, PB1 mutant (K7A D69A), or LIR mutant (D337 338 339A) were collected for immunoblot analysis (n = 5 experiments). (C-E) BUMPT cells stably expressing GFP-SQSTM1/p62 WT, PB1 mutant (K7A D69A), or LIR mutant (D337 338 339A) were incubated for 1 day with or without 5 ng/ml TGFB1 in serum-free DMEM for morphological and immunoblot analyses. (C) Representative fluorescence images showing the formation of GFP-SQSTM1/p62 aggregates. Scale bar: 200 µm. The boxed areas were enlarged and placed on the right side. (D) immunoblots of p-MAPK1/ERK2-MAPK3/ERK1 (Thr202/Tyr204) and MAPK1/ERK2-MAPK3/ERK1. (E) densitometry of p-MAPK1/ERK2-MAPK3/ERK1 (Thr202/Tyr204) immunoblot (n = 5 experiments). Data in (B) and (E) are presented as mean ± SEM and two-way ANOVA with multiple comparisons was used for statistics.