Fig. 3.
TUDCA regulated mitochondrial fission via increased mitochondrial PrPCamounts in CKD-hMSCs. (A) Representative TEM images (n = 3 per group) after incubation of Healthy-hMSCs with or without si-PRNP or after treatment of TUDCA-pretreated CKD-hMSCs with si-PRNP. Scale bars are shown enlarged on the left. Scale bars: 500 nm (n = 3 sample/group). (B) Quantitative analyses of morphometric data from TEM images. Values represent the mean ± SEM. *p < 0.05, and **p < 0.01 vs. untreated Healthy-hMSCs, #p < 0.05 and ##p < 0.01 vs. si-PRNP-treated Healthy-hMSCs, $p < 0.05 and $$p < 0.01 vs. CKD-hMSCs, &p < 0.05 and &&p < 0.01 vs. TUDCA-treated CKD-hMSCs pretreated with si-PRNP. (One-way ANOVA, using Tukey's post-hoc test). (C) Percentages of abnormal mitochondria, which were swollen with evidence of severely disrupted cristae throughout a mitochondrion) obtained from TEM images. Values represent the mean ± SEM. *p < 0.05, and **p < 0.01 vs. untreated Healthy-hMSCs, #p < 0.05 and ##p < 0.01 vs. si-PRNP-treated Healthy-hMSCs, $p < 0.05 and $$p < 0.01 vs. CKD-hMSCs, &p < 0.05 and &&p < 0.01 vs. TUDCA-treated CKD-hMSCs pretreated with si-PRNP. (One-way ANOVA, using Tukey's post-hoc test). (D) Western blot analysis quantified the expression of p-DRP1, MFN1, and OPA1 in Healthy-hMSCs, treatment of CKD-hMSCs with or without TUDCA, and after treatment of TUDCA-pretreated CKD-hMSCs with si-PRNP (n = 3 sample/group). (E) The expression levels were determined by densitometry relative of VACD1. Values represent the mean ± SEM. *p < 0.05 and **p < 0.01 vs. Healthy-hMSCs, #p < 0.05 and ##p < 0.01 vs. CKD-hMSCs, $p < 0.05 and $$p < 0.01 vs. TUDCA-treated CKD-hMSCs pretreated with si-PRNP. (One-way ANOVA, using Tukey's post-hoc test). (F) Western blot analysis quantified the expression of P62, and LC3B in Healthy-hMSCs, treatment CKD-hMSCs with or without TUDCA, and after treatment of TUDCA-pretreated CKD-hMSCs with si-PRNP. (G) The expression levels were determined by densitometry relative to VACD1. Values represent the mean ± SEM. *p < 0.05 and **p < 0.01 vs. Healthy-hMSCs, #p < 0.05 and ##p < 0.01 vs. CKD-hMSCs, $p < 0.05 and $$p < 0.01 vs. TUDCA-treated CKD-hMSCs pretreated with si-PRNP. (One-way ANOVA, using Tukey's post-hoc test).