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. 2022 May 18;19(1):256–277. doi: 10.1080/15548627.2022.2072054

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Figure 2. — Tubular autophagy deficiency in iRT-atg7-KO mice inhibits interstitial fibrosis during post-ischemic kidney repair. (A) Top panel: genomic DNA was extracted from WT (n = 6) and iRT-atg7-KO (n = 6) kidneys at the indicated time points after doxycycline treatment for PCR detection of Atg7 flox and deletion alleles. Other panels: tissue lysates were collected from WT and iRT-atg7-KO kidneys for immunoblots of ATG7, LC3B, ATG5 (ATG12 conjugated), and SQSTM1/p62. (B-F) 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 at the indicated time points for histology, RT-qPCR and immunofluorescence or IHC. (B) Masson’s trichrome staining. Scale bar: 50 µm. (C) Quantification of collagen-stained areas (UI30R 2 w: WT n = 6, KO n = 6; UI30R 4 w: WT n = 6, KO n = 7). (D) RT-qPCR assay of Fn1, Col1a1, Col4a1, and Acta2 mRNA (sham: WT n = 4 or 5, KO n = 4 or 5; UI30R 4 w: WT n = 7, KO n = 9). (E and F) Immunofluorescence or IHC of FN1, COL1A1, COL4A1 and ACTA2 (WT n = 7, KO n = 7). Scale bar: 100 µm. Data in (C and D) are presented as mean ± SD. Two-way ANOVA with multiple comparisons was used for statistics.