Figure 6.

Antioxidant treatment induces rescue of hematopoietic phenotypes in AK2 zebrafish mutants. (A) Embryos from an incross of heterozygous ak2^L124P mutants were treated with different doses of NAC (10, 20, or 50 µM) until 5 dpf when the expression of different markers was assessed by WISH. (left) Quantification of rescue induced by NAC treatment of ak2^L124P missense mutants. Data are presented as the percentage of abnormal expression of each marker by WISH analysis at the indicated concentrations. The number of embryos analyzed is shown above each column. All observed differences compared with the untreated embryos are significant (P < 0.005, Z-test). (right) Representative stereomicroscopy images of tail regions of WT siblings and ak2^L124P/L124P mutants untreated or treated with different doses of NAC. Black arrowheads indicate hmox1a-positive cells in CHT regions (boxed insets). (B) Lateral views of WT siblings and ak2^del2/del2 mutants showing c-myb expression (WISH) in untreated and treated with 50 µM NAC embryos at 5 dpf. Red and black insets show thymus and kidney regions, respectively; white arrows mark insulin expression (positive control). (C) Lateral views of rag 1 expression (WISH) in thymic region of WT siblings, ak2^del2/del2 mutants untreated and treated with increasing concentrations of NAC at 5 dpf. Black circles show thymic region in each sample. (A [right], B, and C) For each sample, at least 25 embryos have been analyzed. Bars, 100 µm. (D and E) Quantitative analysis of the effect of antioxidant treatment on c-myb (D) and rag1 (E) expression at 5 dpf in embryos from an incross of heterozygous ak2^del2 mutants. Data are presented as the percentage of ak2^del2/del2 null embryos. Numbers above columns indicate the total number of ak2^del2/del2-null mutants found in each group of genotyped embryos. Data (A–E) are representative of at least two independent experiments.