2.8 Protein isolation and Western blot
Proteins were extracted from cells and tissues (30 mg) with radioimmunoprecipitation assay (RIPA) buffer, denatured with loading buffer, resolved with 10% SDS-PAGE gels, electroblotted onto NC membranes, and then blocked with 5% nonfat milk. The membranes were incubated with different primary antibodies, followed by incubation with the relative secondary antibodies, and the blots were analyzed by ImageJ.