Fig. 3

MDM2 ubiquitinates and modifies AGPS and promotes its degradation. a Western blot of AGPS expression after ectopic transfection with different E3 or DUBs. b qPCR of the AGPS expression after ectopic transfection with MDM2. c AGPS expression after MDM2 knockdown. n.s. no significance. d Western blot of the AGPS expression after ectopic transfection with different doses of MDM2. e Western blot of the AGPS expression after ectopic transfection with MDM2 and treatment with or without MG132 (20 μM) for 8 h. f Western blot analysis of AGPS protein expression in PC-3 cells after AGPS knockdown followed by treatment with cycloheximide (CHX) (50 μg/mL) for 0-, 4-, 8-, 12-, 16-, or 20- h. g The protein bands were quantified and normalized to the band intensity at the 0-h time point. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. h Ubiquitination experiment on the changes in AGPS ubiquitination levels after ectopic transfection with MDM2 and treatment with MG132 (20 μM) for 8 h. i, j Ectopia and endogenic MDM2 and AGPS binding. k Ubiquitination experiment on the changes in AGPS ubiquitination levels after ectopic transfection with MDM2 and different lysis mutant-type of UB