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. 2013 Jan 15;126(2):484–496. doi: 10.1242/jcs.113035

Fig. 2.

Fig. 2.

Partial silencing of DPAGT1 inhibits canonical Wnt activity. (A) Quantitative PCR of DPAGT1 transcript levels in non-silenced (NS) or DPAGT1 silenced (S) cells (***P<0.001). (B) Immunoblot of GPT in NS and S cells. Bar graph: fold change in GPT levels after normalization to actin. (C) Immunoblot of E-cadherin from NS and S cells. (D) Immunoblot of β-catenin and ABC levels in NS and S cells. Bar graphs: fold change in β-catenin and ABC levels after normalization to GAPDH. (E) Immunoblot of γ-catenin expression in NS and S cells. (F) Immunoblot of pGSK-3β and GSK-3β protein levels in NS and S cells. Bar graph: fold change in pGSK3β/GSK3β levels after normalization to GAPDH. (G) Luciferase reporter activity from the FOP-DPAGT1 vector in NS and S cells (***P<0.001). (H) Luciferase reporter activity from the TOP-Flash vector in NS and S cells (***P<0.001). (I) ChIP analyses of β-catenin and Tcf at the DPAGT1 promoter in NS and S cells after normalization to the IgG control (**P<0.005). (J) ChIP analyses of γ-catenin and Tcf at the DPAGT1 promoter in NS and S cells after normalization to the IgG control (**P<0.005). (K) Immunofluorescence localization of β-catenin, ABC, γ-catenin and LRP5/6, in NS and S cells, counterstained for F-actin and nuclei. Scale bars, 10 µm.