Figure 6: p53 enhances 14-3-3γ ubiquitination. H358 cells were cotransfected with Flag-tagged 14-3-3γ expression vector and His-tagged ubiquitin expression vector. After 16 h, cells were infected with Ad-GFP, Adp53 (wt) and Ad-p53 (mt) virus respectively for 32 h and then harvested. Immunoprecipitation was performed using Anti-Flag M2 affinity gel. The beads were washed successively and ubiquitin binding to 14-3-3γ was detected by SDS-PAGE and immunoblotting using the anti-ubiquitin antibody. 14-3-3γ expression and loading control were also evaluated by anti-Flag and β-actin antibodies, respectively.