Immunoprecipitation and Western blot analysis

Immunoprecipitation was performed as previously described⁵⁰. In brief, the total cell lysates were collected and incubated with corresponding antibodies (HA, H9658, Sigma,1:200; MYC, 2276 S, Cell Signaling Technology,1:250) and protein A/G beads at 4 °C overnight. Immunoprecipitants were separated by SDS–PAGE and visualized by ECL Plus. For western blot, tissue or cell lysates were separated by SDS-PAGE and blotted on PVDF (polyvinylidene difluoride) membranes (Millipore). Then the membranes were incubated with corresponding primary antibody (GCA, PA5-77127, Invitrogen, 1:1000; p-AKT, CST9271s, Cell Signaling Technology,1:1000; AKT, CST9272s, Cell Signaling Technology,1:1000; PAK1, 2602 T, Cell Signaling Technology,1:1000; Phospho-PAK1, 2601 T, Cell Signaling Technology,1:1000; Phospho-NF-κB p65, CST3033S, Cell Signaling Technology,1:1000; NF-κB p65, CST8242S, Cell Signaling Technology, 1:1000; PHB2, sc-133094, Santa Cruz,1:1000) at 4 °C overnight. And specific proteins were visualized by ECL Plus.