Fig. 6: OPN promotes nuclear translocation of β-Catenin by phosphorylating it at Ser675 and activates Wnt signaling pathway. | Cell Death & Disease

Fig. 6: OPN promotes nuclear translocation of β-Catenin by phosphorylating it at Ser675 and activates Wnt signaling pathway.

From: Osteopontin promotes metastasis of intrahepatic cholangiocarcinoma through recruiting MAPK1 and mediating Ser675 phosphorylation of β-Catenin

Fig. 6

a The activity of the TOP/FOP reporter plasmid was analyzed by luciferase reporter assays in ICC cell lines with OPN up- or down-regulation. b β-Catenin could restore the activity of TOP/FOP reporter plasmid induced by OPN knockdown. c Immunofluorescence images of RBE cells with empty vector or OPN over-expressed (left). OPN, β-Catenin, and cell nuclei (DAPI) were exhibited as red, green, and blue, respectively. Cell lysates were subjected to the nuclear and cytosol fractionation, and β-Catenin expression was analyzed by western blot (right). d OPN was stably over-expressed in RBE cells and cell lysates were subjected to Co-IP; the phosphorylation of β-Catenin at S675, S552, Y654, and Y142 was analyzed by western blot. e The activity of kinases related to Wnt signaling activation was evaluated in RBE cells, which stably expressed empty vector or OPN and pretreated with U0126 (10 μM), EHop-016 (10 μM), SP600125 (20 μM), and LY290042 (20 μM) for 24 h. f Co-IP analysis of phosphorylated β-Catenin at Ser675 and OPN-β-Catenin complex was determined by western blot in RBE cells with Flag-β-Catenin and HA-OPN overexpression and pretreated with U0126, EHop-016, SP600125, and LY290042 for 24 h

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