Fig. 2 S3

HSPCapoptosis but no gross morphological and vascular defects in in dnmt3bb.1 morphants.

(a,b) Immunohistochemical staining of the trunks of 48 hpf control (a) and dnmt3bb.1 (b) morpholino-injected Tg(cmyb:gfp) transgenic zebrafish with anti-GFP (green) and anti-caspase3 (red) antibodies, showing increased numbers of caspase3-positive HSPC in dnmt3bb.1-deficient animals. (c) Quantitation of the percentage of GFP (cmyb) positive HSPCs that are also caspase3-positive in 48 hpf control or dnmt3bb.1 morpholino-injected Tg(cmyb:gfp) transgenic zebrafish. Columns show mean ± SEM, and statistical significance calculated by paired t-test is shown (p = 0.0012). (d,e) Transmitted light images of whole 36 hpf control (d) and dnmt3bb.1 (e) morpholino-injected zebrafish, showing absence of developmental delay or gross morphological abnormalities in dnmt3bb.1 morphants. (f,g) Confocal images of trunk EGFP fluorescence in 36 hpf control (f) or dnmt3bb.1 (g) morpholino-injected Tg(fli1a:egfp)^y1 transgenic zebrafish, showing normal trunk vessel patterning in dnmt3bb.1 morphants. (h,i) Whole mount in situ hybridization of the trunks of 36 hpf control (h) or dnmt3bb.1 (i) morpholino-injected zebrafish probed for vecdn, showing normal expression in dnmt3bb.1 morphants. All images are lateral views, rostral to the left. Scale bars 25 µm in b, 100 µm in e and 50 µm in g,i.