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. 2016 Jul 22;7(33):53289–53298. doi: 10.18632/oncotarget.10795

Figure 5. PDIA6 inhibits ubiquitination and proteasomal degradation of β-catenin.

Figure 5

(A) HeLa cells transfected with pCMV-sport6-PDIA6 or pCMV-sport6 were treated with the proteasome inhibitor MG-132 for 6 h. β-TrCP was not treated with MG-132. Total protein was extracted for Western blotting analysis. (B) The quantitative analysis of these proteins was normalized to β-actin. *p < 0.05. (C) HeLa cells transfected with plasmid were treated with CHX to block protein synthesis or vehicle DMSO for western blotting analysis at the indicated time points. PDIA6 accelerated degradation of β-TrCP compared to negative control cells. (D) DMSO treatment was a positive control to eliminate the interference of DMSO in the same experimental condition. (E) The quantitative analysis of normalized β-TrCP levels. (F) The overexpression of PDIA6 decreased β-catenin ubiquitination relative to the negative control. HeLa cells were transfected and immunoprecipitated with anti-β-catenin antibody, followed by immunoblotting with an anti-ubiquitin antibody.