Fig. 3: The anti-fibrotic effects of 1, 5-AG in UUO model.

A Picture of left kidneys of mice with different treatments (n = 3). B The renal index (mg/g) (n = 6). ***p = 0.0008 for B: Sham vs. UUO, p = 0.3372 for B: Sham vs. 1, 5-AG, ^#p = 0.0221 for B: UUO vs. 1, 5-AG. C Western blots show fibrotic protein expression in kidneys from all groups. D Quantitative analysis of fibrotic protein (n=3). ***p = 0.0002 for FN: Sham vs. UUO, p = 0.0637 for FN: Sham vs. 1, 5-AG, ^##p = 0.0017 for FN: UUO vs. 1, 5-AG; **p = 0.0024 for COL-1: Sham vs. UUO, p = 0.9068 for COL-1: Sham vs. 1, 5-AG, ^##p = 0.0043 for COL-1: UUO vs. 1, 5-AG; ***p < 0.0001 for α-SMA: Sham vs. UUO, *p = 0.0170 for α-SMA: Sham vs. 1, 5-AG, ^##p = 0.0011 for α-SMA: UUO vs. 1, 5-AG. E Biochemical parameters including BUN, Scr, TC, TG in each of mice (n = 5). ***p = 0.0002 for BUN: Sham vs. UUO, *p = 0.0319 for BUN: Sham vs. 1, 5-AG, ^#p = 0.0361 for BUN: UUO vs. 1, 5-AG; *p = 0.0146 for SCR: Sham vs. UUO, p = 0.2637 for SCR: Sham vs. 1, 5-AG, ^#p = 0.0309 for SCR: UUO vs. 1, 5-AG, Comparison in SCR were performed with a two-tailed Student’s t test; *p = 0.0159 for TC: Sham vs. UUO, p = 0.9351 for TC: Sham vs. 1, 5-AG, ^#p = 0.0429 for TC: UUO vs. 1, 5-AG; **p = 0.0038 for TG: Sham vs. UUO, p = 0.9255 for TG: Sham vs. 1, 5-AG, ^#p = 0.0107 for TG: UUO vs. 1, 5-AG. F Representative photomicrographs of the H&E staining and Masson’s trichrome staining from left kidneys of Sham, UUO, and 1, 5-AG-treated UUO mice (H&E and Masson’s staining; scale bar, 100 μm; magnification, ×200). G Bar graphs depict renal injury scores and renal interstitial fibrosis scores based on H&E staining or Masson’s trichrome staining (n = 3). **p = 0.0012 for injury scores: Sham vs. UUO, *p = 0.0423 for injury scores: Sham vs. 1, 5-AG, ^#p = 0.0332 for injury scores: UUO vs. 1, 5-AG; ***p = 0.0003 for fibrosis scores: Sham vs. UUO, *p = 0.0139 for fibrosis scores: Sham vs. 1, 5-AG, ^##p = 0.0079 for fibrosis scores: UUO vs. 1, 5-AG. H Kidney expression of TGF-β/Smad signaling pathway from Sham, UUO and 1, 5-AG-treated UUO mice, assayed by Western blot. (I) Quantitative analysis of Fig. 3H (n = 3). **p = 0.0031 for TGF-β: Sham vs. UUO, p = 0.4396 for TGF-β: Sham vs. 1, 5-AG, ^#p = 0.0138 for TGF-β: UUO vs. 1, 5-AG; ***p = 0.0002 for SMAD2: Sham vs. UUO, **p = 0.0053 for SMAD2: Sham vs. 1, 5-AG, ^#p = 0.0126 for SMAD2: UUO vs. 1, 5-AG; ***p = 0.0002 for SMAD3: Sham vs. UUO, p = 0.3792 for SMAD3: Sham vs. 1, 5-AG, ^###p = 0.0006 for SMAD3: UUO vs. 1, 5-AG. J Representative Western blot of Nrf2, Keap1, 12-LOX, Rac-1. (K) Quantitative analysis of Fig. 3J (n = 3). *p = 0.0156 for NRF2: Sham vs. UUO, p = 0.4461 for NRF2: Sham vs. 1, 5-AG, ^#p = 0.0331 for NRF2: UUO vs. 1, 5-AG; ***p  < 0.0001 for KEAP1: Sham vs. UUO, **p = 0.0019 for KEAP1: Sham vs. 1, 5-AG, ^###p = 0.0007 for KEAP1: UUO vs. 1, 5-AG; ***p  < 0.0001 for 12-LOX: Sham vs. UUO, p = 0.9980 for 12-LOX: Sham vs. 1, 5-AG, ^###p  < 0.0001 for 12-LOX: UUO vs. 1, 5-AG; ***p  < 0.0001 for RAC1: Sham vs. UUO, p = 0.1273 for RAC1: Sham vs. 1, 5-AG, ^###p  < 0.0001 for RAC1: UUO vs. 1, 5-AG. Data are presented as mean ± SD. Comparisons in B, D, E, G, I and K were compared using One-Way ANOVA followed by Sidak’s multiple comparisons test. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 (compared with sham group), ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 ⁽compared with UUO group). Individual data points are independent biological replicates unless otherwise stated.