FIGURE 8.

ZNF213 destabilizes YAP possibly through K48‐linked poly‐ubiquitination. A, ZNF213 prohibits YAP degradation through its full length. HEK293 cells were transfected with 2 µg GFP‐ZNF213 full length or mutant plasmids (ΔZF, ΔKRAB‐A+ΔZF, ΔLeR/SCAN, ΔLeR/SCAN+ΔKRAB‐A). About 24h later, cells were treated with 10 µM MG132 for 6 h. Cells were harvested directly and used for the analysis of western blotting. B, ZNF213 increases the overall poly‐ubiquitination of YAP. HEK293 cells were transfected with 1 mg Myc‐ZNF213 or Myc‐tag plasmid. About 24h later, cells were transfected with 1 mg HA‐Ub plasmid. Another 24h later, the cell extracts were immunoprecipitated with HA antibody. The poly‐ubiquitinated YAP was detected through the analysis of western blotting. C, ZNF213 increases K48‐linked poly‐ubiquitination of YAP. HEK293 cells were transfected with 1 mg Myc‐ZNF213 or Myc‐tag plasmid, together with 1 mg HA‐K48 Ubi plasmid. The cell extracts were immunoprecipitated with HA antibody. The K48 specific poly‐ubiquitinated YAP was detected through the analysis of western blotting. D, ZNF213 decreases K63‐linked poly‐ubiquitination of YAP. HEK293 cells were transfected with 1 mg Myc‐ZNF213 or Myc‐tag plasmid, together with 1 mg HA‐K63 Ubi plasmid. The cell extracts were immunoprecipitated with HA antibody. The K63 specific poly‐ubiquitinated YAP was detected through the analysis of western blotting