Fig. 3

Fig. 3 p53-dependentproapoptotic signaling mediates the death of immature T lymphocytes.

A Comparison of the expression of critical p53 signaling genes and cell cycle genes. B Relative expression of tp53, the tp53 downstream genes 113p53, mdm2 and p21, puma, bax, bik and the cell cycle gene ccng1. Besides the expression of tp53, all of the genes were upregulated in mcm5^−/− embryos. As shown by RT‒qPCR, the experiments were repeated at least 3 times, and the value was normalized to that of β-actin. C The expression of tp53 was evaluated by WISH and Western blotting for siblings, mcm5^−/− embryos and siblings treated with heat-shock. The right columns show the relative gray level for Western blotting. The mRNA level of tp53 was not upregulated in mcm5^−/− embryos, but the protein level was greatly increased. D TUNEL staining was evaluated for Coro1a:GFP-labeled cells at 3.5 dpf in siblings (n = 7), mcm5^−/− (n = 7) and mcm5^−/− embryos injected with p53MO (n = 7). E On 4 dpf, the number of Coro1a: GFP-labeled cells in siblings (e1, n = 38), mcm5^−/−embryos (e2, n = 29), embryos injected with p53 MO (e3, n = 34) and mcm5^−/−embryos injected with p53 MO (e4, n = 24) was examined. The number of Coro1a: GFP-labeled cells in mcm5^−/−embryos was partially restored by injecting p53MO (e5). F The expression of rag2 was examined in siblings (f1, n = 20), mcm5^−/−embryos (f2, n = 15), embryos injected with p53 MO (f3, n = 17) and mcm5^−/−embryos injected with p53 MO (f4, n = 13). The expression of rag2 in mcm5^−/−embryos was partially restored by injecting p53MO (f4-f5). Scale bars, 40 μm. For (B, C), e5 and f5, the data were presented as means ± SD; The P-values (t-test; two-tailed); NS, not significant. “*” P < 0.05, “**” P < 0.01, “***” P < 0.001.