Journal: OncoTargets and therapy

Article Title: Knockdown of RNF6 inhibits gastric cancer cell growth by suppressing STAT3 signaling

doi: 10.2147/OTT.S174846

Figure Lengend Snippet: 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.

Article Snippet: Anti-PARP, cyclin D1, p-STAT3, STAT3, SHP-1, MCL1, Bcl-2 and GAPDH antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA; dilutions: 1:1,000).

Techniques: Infection, Western Blot, Quantitative RT-PCR, Transfection, Immunoprecipitation, Plasmid Preparation, Real-time Polymerase Chain Reaction