Figure 3.
High levels of ROS in TCS embryos and prevention of craniofacial malformation through the pharmacological suppression of ROS. (a) Level of ROS measured in control (STD), TRA and TRA+ATGTp53 48 hpf embryos. Values are expressed as the mean±S.E.M. (n=42). **P<0.01 versus STD; NS= non-significant P>0.05 (t-test) AU: arbitrary units. (b) Frequencies of Normal, Mild and Severe phenotypes (see Results section) in controls (STD, n=51), TRA (n=79) and TRA-larvae incubated in the presence of NAC (TRA+NAC, n=52). **P<0.01 ***P<0.001 versus STD; ##P<0.01 versus TRA (chi-square test). (c) Acridine Orange staining in vivo performed in 24 hpf embryos (see Figure 2c for representative photos of normal (+), intermediate (++) or extensive (+++) cephalic cell death). The bar graph shows the distribution of each phenotype among the groups (STD, TRA and TRA+NAC embryos). n=25 embryos. ***P<0.001 versus STD; NS=non-significant P>0.05 (chi-square test). (d) Western blotting showing the Tp53 abundance in STD, TRA and TRA+NAC 24 hpf embryos. Molecular weight markers are shown at the right. Actin was analyzed as a loading control. The graph shows the densitometric quantification of western blotting band (normalization of Tp53 signal with respect to Actin). Bars represent mean of relative abundance and S.E.M., n=3, *P<0.05 NS,non-significant P>0.05 (Mann–Whitney test)