Fig. 4

PHF5A stabilized VEGFA protein via interacting with MDM2. (A) The protein level of potential PHF5A targets were detected by WB analysis. (B) The interaction between PHF5A (Flag) and VEGFA was assessed by Co-IP analysis. (C) The E3 ligase of VEGFA was predicted using ubibrowser website. (D) The endogenous interaction between PHF5A and MDM2 was revealed via Co-IP experiment. (E) The VEGFA protein half-life was decreased upon PHF5A depletion or MDM2 overexpression. The cells were treated with 50 µg /mL CHX at the indicated time. (F) The addition of MG132 attenuated the VEGFA protein degradation induced by PHF5A depletion or MDM2 overexpression. The transfected Eca-109 cells were treated with 10 µM MG132 for 24 h. (G) The endogenous VEGFA protein ubiquitination was promoted by PHF5A silencing and MDM2 overexpressing. The transfected Eca-109 cells were treated with 20 µM MG132 for 6 h. GAPDH served as an internal control