Fig. 2
Short term phenotype of Sirt1 -/- mutants.
Four-days post fertilization embryos were used to obtain representative images of short term phenotypes of wildtype and Sirt1-/- mutants. (A) Phenotypes of wildtype and Sirt1-/- zebrafish embryos. Note there are no differences in appearance between wildtype and Sirt1-/- mutant embryos. (B) The embryo survival curves of wildtype and Sirt1-/- mutants after treatment with 0.5 or 1 mM tBOOH. Note that wildtype embryos died earlier than Sirt1-/- mutants. (C) ROS levels in 0.5 mM tBOOH-treated wildtype and Sirt1-/- embryos were examined by H2DCFDA fluorescence. (D) Detection of apoptosis in 0.5 mM tBOOH-treated wildtype and Sirt1-/- mutant embryos by TdT-mediated dUTP nick end labeling (TUNEL). Note that Sirt1-/- mutants showed higher levels of ROS and TUNEL than wildtype embryos. The effect of NAC treatment on ROS content and apoptosis incidence was studied by H2DCFDA fluorescence (E) and the TUNEL assay (F). Embryos were exposed to treatment with NAC treatment for 24 h. Results showed that TUNEL-positive cells increased in both, tBOOH-treated wildtype and Sirt1-/-groups; furthermore, NAC treatment was effective for both groups.