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Fig. 5. TNF{alpha} activates p53 and induces its phosphorylation at serine 18 through p38 kinase. (A) Abundance and status of phosphorylated p53 at serine 18, p53 and p21 in TNF{alpha}-treated MEFs. Serum-arrested MEFs were treated with TNF{alpha} for the indicated time. Total protein lysates were prepared and analysed by gel electrophoresis followed by immuno-blotting analysis using antibodies to p53 phosphorylated at serine 18 (pp53/Ser18), p53 and p21. Each lane contains equal amounts of loaded proteins (40 µg). (B) Abundance and status of phosphorylated p53 at serine 18, p53 and p21 in TNF{alpha}-treated MEFs prealably treated with SB 203580. Serum-arrested cells were pre-treated with SB 203580, followed by a treatment with TNF{alpha} for the indicated time. Lysates were analysed as in (A). (C) Abundance and status of phosphorylated p53 at serine 18, p53 and p21 in TNF{alpha}-treated MEFs expressing Cdc42-N7. Cells were transfected with GFP-tagged Cdc42-N17 before serum-starvation, followed by a treatment with TNF{alpha} for the indicated time. Lysates were analysed as in (A).