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

Figure 2.

Figure 2.

Autophagy deficiency inhibits EGR1 in cultured mouse renal proximal tubular cells during TGFB1 treatment. (A and B) subconfluent BUMPT cells were exposed to 5 ng/ml TGFB1 in serum-free DMEM for 2 days alone or with 5 µM CQ or 1 mM 3-MA. Control cells were kept in serum-free medium without TGFB1. Cells were collected for immunoblot of LC3B and SQSTM1/p62 (A) and RT-qPCR of Egr1 mRNA (B) (n = 6 experiments). (C-E) subconfluent WT and atg7 KO cells were exposed to 5 ng/ml TGFB1 in serum-free DMEM for 2 days. Control cells were kept in serum-free medium without TGFB1. (C) RT-qPCR of Egr1 mRNA (n = 6 experiments). (D) immunoblot of EGR1, p-SMAD2 (Ser465/467), SMAD2-SMAD3, p-NFKB RELA/p65 (Ser536), NFKB RELA/p65, p-TRP53/p53 (Ser15), TRP53/p53 and LC3B. (E) densitometry of EGR1 immunoblot (n = 6 experiments). Data in (B), (C) and (E) are presented as mean ± SEM. For statistics, one-way ANOVA with multiple comparisons was used for (B). Two-way ANOVA with multiple comparisons was used for (C) and (E).