TRP53 dependent expression of Bax in cultured blastocysts.(A) Western blotting analysis using anti-Bax antibody of blastocysts collected from the uterus on day 4 (Fresh) or of embryos produced by IVF and cultured for 96 h (113 h post hCG). The embryos were extracted as groups of 20, 40 or 80 embryos and then subjected to Western blotting analysis. Approximately 500 F9 cells were also analyzed as positive controls. The blots shown are representative of 3 experiments. (B) The localization of Bax by indirect immunofluorescence (using same antibody as in Western blot analysis) with blastocysts collected from the uterus (Fresh) or those fertilized within the reproductive tract and the cultured for 96 h (Cultured) (113 h post hCG). Controls were subjected to IgG instead of primary antibody (IgG). The cultures were performed with Trp53+/+ and Trp53-/- embryos. Whole section images were deconvoluted and converted to pseudocolor representation of staining intensity. The sections shown are representative of 3 replicate experiments with a minimum of 8 embryos per treatment per replicate.