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Expression vectors encoding wild-type p53, p53F19A, p53F270A and p53F19A/F270A (1 μg of DNA) were transfected into H1299 cells, and the expressed proteins were examined for changes in their steady-state levels of p53 with the amount of ubiquitination products determined by immunoblotting with anti-p53 antibody (DO-1, top panel), anti-MDM2 antibody (middle panel) or anti-p21 antibody (bottom panel) as indicated.
