Immunoprecipitation and Western blot analysis. a. Immunoprecipitation: lung extract from E15.5 wild-type embryos was used for immunoprecipitation using Smad2 and p53 antibodies. The Western blots were probed with pSmad2, Fbox11 and p53 antibodies. b. Western blot analysis: protein lysates from wild-type, heterozygote and homozygote E15.5 palates. Equal amounts of protein were subjected to tris-acetate PAGE, transferred and probed with pSmad2, Fbox11, p53 antibodies and actin antibody for loading control. c. Graphical representation of p53 levels normalized against actin in wild-type, heterozygote and homozygote E15.5 palates, showing approximately threefold reduction of p53 in homozygote palate compared with wild type.