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. 2016 Mar 3;7(16):21939–21951. doi: 10.18632/oncotarget.7877

Figure 4. Prp19 inhibits the ubiquitin/proteasome-dependent degradation of Twist1 in HCC cells.

Figure 4

A, B.* Stable Huh7 cells mis-expressing Prp19 were treated with 100μg/ml cycloheximide (CHX) for indicated time, and then Twist1 expression were analysed. C. Huh7 cells were transfected with indicated siRNAs. After 48h transfection, Huh7 cells were treated sequentially with 50μg/ml CHX for 1h, 20μM MG132 for another 3h, and followed by western blot. D. Huh7 cells were transfected with indicated siRNAs or plasmids for 48h, and then treated with 20μM MG132 for another 6h. The ubiquitination of Twist1 was assessed by immunoprecipitation (IP) and immunoblot (IB). E. 293T cells (left panel) and Huh7 cells (right panel) were transfected with indicated plasmids, and then WT-Twist1 and Ser68A-Twist1 were detected using antibody against Flag. F.* Huh7 cells were transfected with indicated plasmids and treated with 100 μg/ml CHX for indicated time points, followed by western blot. *Relative densitometric values were detected and presented.