Article Title: MDM2 mediates p73 ubiquitination: a new molecular mechanism for suppression of p73 function
Figure Lengend Snippet: Overexpression of Mdm2 promotes p73 degradation in Mdm2-null MEFs A. Mdm2 null MEFs were transfected with plasmids expressing p73α or an empty vector with Myc-tagged Itch, along with a GFP expressing plasmid (pEGFP) and analyzed by western blotting with a p73-specific antibody (H-79), a Myc-specific antibody for Myc-Itch, a GFP-specific antibody (B-2), and actin as a loading control. B. The same procedure as (A) was used, except that Mdm2 null MEFs were cotransfected with plasmids expressing GFP, p73α along with Itch and Mdm2. C. and D. Mdm2-null MEFs were transfected with an Itch-specific siRNA or a control siRNA. Thirty hours later, the cells were transfected with plasmids expressing GFP, Mdm2 and analyzed by western blot using anti-p73, anti-Itch, anti-p53 (Pab421), anti-GFP (B-2), and anti-Mdm2 (MD-219) antibodies as indicated. E. Mdm2-null MEFs were transfected with plasmids expressing GFP and increased amounts of Mdm2 and analyzed by western blotting with p73-specific (H-79), GFP-specific (B-2) and Mdm2-specific (MD-219) antibodies.
Article Snippet: siRNA experiments For siRNA experiments, HEK293 cells were transfected with siRNA using Lipofectamine 2000 (Invitrogen).
Techniques: Over Expression, Transfection, Expressing, Plasmid Preparation, Western Blot