Figure 5.

RNF6 regulates the stability of SHP-1 by inducing its polyubiquitination.

Notes: (A, B) NCI-N87 cells were infected with shNC, shRNF6#1 or shRNF6#2 for 3 days. Then, cells were prepared for immunoblotting and qRT-PCR as indicated. (C, D) NCI-N87 cells were transfected with increased concentrations of Myc-RNF6 plasmids. Three days later, cells were prepared for immunoblotting and qRT-PCR as indicated. (E) NCI-N87 cells were transfected with EV or Myc-RNF6 plasmids for 3 days, and then cells were prepared for immunoprecipitation with normal IgG or SHP-1 antibody, followed by immunoblotting analysis against Ub and SHP-1.

Abbreviations: EV, empty vector; n.s., nonsense; qRT-PCR, quantitative real-time PCR.