Ectopic expression of JAZ up-regulates expression of p21 and BAX, induces Rb dephosphorylation, and represses cyclin A expression. Cotransfection of JAZ-GFP or FLAG-JAZ in p53^+/+ and p53^–/– MEFs (A,B,D) and NIH3T3 cells (C) was performed as described in Figure 5. JAZ-GFP– or GFP-positive cells were sorted by FACS. Western blot analysis was performed using antibodies against GFP; FLAG; cyclins D, E, and A; p53; p21; p27; BAX; Rb; and/or tubulin (as a loading control). (A) The densitometric quantitation of protein expression was performed. The average ratio of p21 or BAX compared with tubulin is shown. The ratio is set to 1.00 for the GFP-only–expressing p53^+/+ cells. Three independent experiments with similar results were performed, and a representative figure is displayed. G indicates GFP; J-G, JAZ-GFP; V, pcDNA3; F-J, FLAG-JAZ/pcDNA3; p53, p53wt/pCMV; and p53ΔC, p53ΔC(30)/pcDNA3.