Fig. 6

AK2 bound to endogenous CDK4 within cells. a Detection of binding between AK2 and CDK4 within cells as shown using western blot analysis. Proteins of HeLa and MCF-7 cells were separated using 12% SDS-PAGE then were transferred to PVDF membranes. PVDF membranes were sequentially probed with AK2 and anti-V5 tag antibody to detect CDK4, while PVDF membranes probed only with anti-V5 tag antibody served as the NC, while β-actin served as loading control. The original blot image is shown in Additional file 1: Fig S3. b Analysis of interactions between AK2 and CDK4 in cells using co-immunoprecipitation assays. HeLa and MCF-7 cell lysates pre-incubated with AK2 were co-immunoprecipitated with anti-V5 tag antibody then were subjected to western blot analysis to detect CDK4 binding. Lysates without added purified AK2 served as the negative control (NC). Mouse IgG served as NC for probing with anti-V5 tag antibody. Western blots of HeLa and MCF-7 cell lysates served as input control, while β-tubulin served as loading control. The original blot image is shown in Additional file 1: Fig S4