HERS-H1 cells-derived ELVs activated Wnt/β-Catenin signaling. (A) Immunoblots of exosomal (Hsp70, CD63) and cytoplasmatic cell (actin) markers; Wnt3a proteins in ELVs are presented in the panel. (B) Real time RT-PCR showing the upregulated expression of AXIN2 and TCF7 in DPC cells after treatment with ELVs (80 μg/mL). (C) Western blotting revealing the upregulated expression of β-catenin in DPC cells after treatment with ELVs (80 μg/mL), whereas the same marker was significantly downregulated in the ELVs+DKK1 and DKK1 groups. (D) Immunofluorescence staining of β-catenin in DPC cells after treatment with ELVs (80 μg/mL) alone or combined with DKK1. In the control, β-catenin mostly existed in the cytosol of DPC cells, even after addition of DKK1. ELVs induced the transference of β-catenin from the cytosol into the nucleus (white arrow). Accordingly, addition of DKK1 into ELVs could inhibit the transference of β-catenin from the cytosol into the nucleus. (E) Treatment with ELVs (80 μg/mL) upregulated the expression of DMP1 and DSPP, which was attenuated by treatment with DKK1. (DSPP: dentin sialophosphoprotein; DMP1: dentin matrix protein 1). Scale bars are shown. *p < 0.05 vs. Con; #p < 0.05 vs ELVs.