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. 2021 Sep 28;29(3):556–567. doi: 10.1038/s41418-021-00874-0

Fig. 5. TRIM31 was involved in TGF-β1-mediated fibrosis and inflammation in vitro and in Smad and non-Smad signal pathway activation.

Fig. 5

A Representative Collagen I, III and IV, Fibronectin, α-SMA, MMP2, MMP9, TNF-α, IL-6, and IL-1β western blot images in HK2 cells stimulated with TGF-β1 (10 ng/mL) for 12 h or 24 h, after normal control (NC) and si-TRIM31 transfections. B Relative tnf-α, il-6, and il-1β mRNA levels in HK2 cells in the aforementioned six groups. n = 5 per group. C Relative collagen I, III and IV, fibronectin, mmp2, mmp9, α-sma, and trim31 mRNA levels in HK2 cells in aforementioned six groups. n = 5 per group. D Representative collagen I, III and IV, fibronectin, α-SMA, MMP2, MMP9, and Flag Western blot images in HK2 cells stimulated with TGF-β1 for 24 h and transfected with control vector, TRIM31, TRIM31-C53A/56A, or TRIM31-ΔRing expression plasmid. E Relative tnf-α, il-6, il-1β, and trim31 mRNA levels in HK2 cells of eight groups. n = 5 per group. F Representative Western blot images of Smad2, Smad3, P65, ERK, JNK, and P38 phosphorylation levels in TRIM31+/+ and TRIM31−/− mouse kidneys treated with saline or AngII for 42 days. G Representative Western blot images of Smad2, Smad3, P65, ERK, JNK, and P38 phosphorylation levels in HK2 cells stimulated with TGF-β1 (10 ng/mL) for 0.5 h or 1 h after NC and si-TRIM31 transfections. GAPDH was used for normalization. Data were presented as mean ± SEM and normal distributions were tested by Shapiro–Wilk method, which showed that all the data were normally distributed. Two-way ANOVA followed by Sidak post hoc test was used for B and C. Two-way ANOVA followed by Tukey post hoc test was used for E. Adjusted P values were provided in case of multiple groups comparisons. * adjusted P < 0.05, ** adjusted P < 0.01, *** adjusted P < 0.001. Each experiment was repeated independently for a minimum three times.