Loss of asxl1 induces apoptosis in cmyb-expressing embryonic HSPCs. WISH for cmyb was performed at 36 hpf (A) and 3 dpf (B-E) in asxl1+/+, asxl1+/− and asxl1−/− zebrafish embryos. Boxes in panel B are shown at higher magnification in C-E. (F-H) HSPCs (GFP; green) and cells undergoing apoptosis (TUNEL, TMR red, Roche) in the CHT of 48-hpf asxl1+/+, asxl1+/− and asxl1−/− zebrafish embryos in the Tg(cmyb:EGFP) reporter line background were identified by immunofluorescence microscopy following a dual TUNEL/anti-GFP assay. Arrows indicate apoptotic HSPCs with combined EGFP and DMR red fluorescence signals. (I) cmyb:GFP+/TUNEL+ cells in asxl1+/+, asxl1+/− and asxl1−/− were quantified as a percentage of total cmyb:GFP+ cells. Bars represent the mean and s.e.m. for 8-9 embryos each. Unpaired Student’s t-tests and Prism software were used to determine the P-value for each genotypic group compared with control cells.
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