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. 2020 Mar 20;9(3):36. doi: 10.1038/s41389-020-0220-5

Fig. 6. RNF187 associates with YAP and promotes YAP K48-linked poly-ubiquitination.

Fig. 6

a Co-IP assay revealed association between endogenous RNF187 and YAP protein in BT549 cells. BT549 cells were harvested with RIPA lysis buffer. CO-IP was performed using antibody as indicated. b RNF187 depletion decreased the overall poly-ubiquitination of YAP. BT549 cells were transfected with 50 µM siControl or siRNF187. After 24 h, cells were transfected with 1 µg HA-Ub plasmid. After another 24 h, the cell extracts were immunoprecipitated with HA antibody. The poly-ubiquitinated YAP was detected via western blotting analysis. c RNF187 increased the overall poly-ubiquitination of YAP. HEK293 cells were transfected with 0.5 µg Myc-RNF187 or Myc vector. After 24 h, cells were transfected with 1 µg HA-Ub plasmid. After another 24 h, the cell extracts were immunoprecipitated with HA antibody. The poly-ubiquitinated YAP was detected via western blotting analysis. d RNF187 depletion decreased the K48-linked poly-ubiquitination of YAP. BT549 cells were transfected with 50 µM siControl or siRNF187. After 24 h, cells were transfected with 1 µg HA-K48-Ubi plasmid. After another 24 h, the cell extracts were immunoprecipitated with HA antibody. The K48-linked poly-ubiquitinated YAP was detected via western blotting analysis. e RNF187 increases K48-linked poly-ubiquitination of YAP. HEK293 cells were transfected with 0.5 µg Myc-RNF187 or Myc vector, together with 1 µg HA-K48 Ubi plasmid. The cell extracts were immunoprecipitated with HA antibody. The K48 specific poly-ubiquitinated YAP was detected via western blotting analysis.