Fig 9

(A) Tg(4xnfbr:d2EGFP) is a reporter line for Nfat activity. Representative images of gata1 mutants and controls in the Tg(4xnfbr:d2EGFP);Tg(kdrl:nls-mCherry) background at 65hpf. (B) Representative images of valves in 84 hpf Tg(4xnfbr:d2EGFP);Tg(kdrl:nls-mCherry) embryos that have been treated with DMSO or FK506 from 60 hpf. (C) Representative images of 98 hpf valves in Tg(fli1a:gal4;UAS:Kaede) embryos treated with DMSO or FK506 from 60 to 98 hpf. (D) Representative images of embryos treated with DMSO or FK506 from 60 to 98 hpf and EdU from 74 to 98 hpf. (E) Graph showing number of Nfat activated cells in 65 hpf superior AV valves as determined using the Tg(4xnfbr:d2EGFP);Tg(kdrl:nls-mCherry) line. p-Values were calculated using unpaired t test. (F) Graph showing percentage of embryos with thick superior AV valves in embryos treated with DMSO and FK506 from 60 to 98 hpf. Statistical significance was calculated using Fisher exact test. (G) Graph showing number of cells in the superior AVC in embryos treated with DMSO and FK506 from 60 to 98 hpf. Statistical significance was calculated using Student t test. (H) Graph showing the fraction of EdU positive cells in the superior AVC in embryos treated with DMSO or FK506 from 60 to 98 hpf and EdU from 74 hpf to 98 hpf. Statistical significance determined by Student t test. (I) Graph showing the fraction of EdU positive luminal cells and EdU positive abluminal cells in embryos treated with DMSO or FK506 from 60 to 98 hpf and EdU from 74 hpf to 98 hpf. Statistical significance was determined by 2-way ANOVA. AV, atrioventricular valve; AVC, atrioventricular canal; hpf, hours postfertilization; Nfat, nuclear factor of activated T cells.