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. 2021 Oct 5;22(19):10780. doi: 10.3390/ijms221910780

Figure 4.

Figure 4

HLSC-EVs downregulate β-catenin and pro-fibrotic marker α-Sma in mkCF in an in vitro model of AAN. (A) A schematic representation of the transwell setting of the AAN model in vitro. Briefly, mTECs pre-treated with 100 µM AA for 4 h were co-cultured with mkCF in the presence or absence of HLSC-EVs. Post-experimental analysis revealed an up-regulation of the genes Ctnnb1 (β-catenin) (B) and the profibrotic marker α-Sma (C) in mkCF exposed to AA-treated mTECs. Treatment with HLSC-EVs downregulated both genes significantly. Western blot analysis further revealed an up-regulation of active β-catenin (D) and α-SMA (E) at the protein level, which was significantly downregulated by HLSC-EVs. The data represent mean ± SEM of three independent experiments performed in quadruplicate. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. mTEC+AA. A one-way analysis of variance with Bonferroni’s multi-comparison test was performed.