Fig. 4

Abnip3lb qPCR in bnip3lb splice-morphant embryos at 72 hpf (**P = 0.0015) [two-tailed unpaired t test with Welch’s correction: mean ± SEM, n = 30 embryo tails/condition, 3 clutches]. B Quantification of mitophagy positive puncta in HSPCs in the VDA at 48 hpf, delimited by Kdrl:BFP expression, in controls and bnip3lb splice-morphants (*P = 0.0474) [two-tailed unpaired t test with Welch’s correction: mean ± SEM, n = 4]. C Representative confocal image of mitophagy (pink arrows) in the VDA at 48 hpf, marked by Kdrl:BFP expression, with or without bnip3lb knockdown (representative of n = 4/clutch). Scale bar, 50 µM. D Representative WISH images of runx1/cmyb expression in the VDA at 36 hpf in bnip3lb splice morphant embryos versus WT control. Scale bar, 100 µM. E Phenotypic distribution of runx1/cmyb expression scored in embryos from (D) (**P = 0.0010) [Pearson’s chi-square (two-sided): n = 20/condition, 5 clutches]. F Representative WISH image of runx1/cmyb expression in the CHT at 72 hpf following splice-targeted bnip3lb knockdown. Scale bar, 100 µM. G Phenotypic distribution of runx1/cmyb expression scored in embryos from (F) (**P = 0.003) [Pearson’s chi-square (two-sided): n = 22/condition, 3 clutches]. H Frequency of CD41^lo HSPCs in Tg(−6.0itga2b:EGFP) embryos by flow cytometry at 72 hpf after splice-targeted bnip3lb MO knockdown versus controls (*P = 0.0368) [two-tailed unpaired t test with Welch’s correction: mean ± SEM, n = 8/condition, 6 clutches]. I Frequency of CD41^lo HSPCs in Tg(−6.0itga2b:EGFP) embryos by flow cytometry at 72 hpf after ATG-targeted bnip3lb MO knockdown versus controls (*P = 0.0456) [two-tailed unpaired t test with Welch’s correction: mean ± SEM, n = 6/condition, 8 clutches]. J Frequency of CD41^lo HSPCs in Tg(−6.0itga2b:EGFP) embryos by flow cytometry at 72 hpf after CRISPR/CAS9-mediated knockout of bnip3lb (*P = 0.0439) [two-tailed unpaired t test with Welch’s correction: mean ± SEM, n = 12/condition, 6 clutches]. K Representative WISH images of rag1 expression in the thymus at 120 hpf in control and splice-targeted bnip3lb morphants. Scale bar, 100 µM. L Phenotypic distribution of rag1 expression scored in embryos from (K) (*P = 0.0409) [Pearson’s chi-square (two-sided): n = 22/condition, 4 clutches]. M Frequency of Rag2+ lymphoid progenitor cells in Tg(rag2:GFP) embryos by flow cytometry at 120 hpf after bnip3lb knockdown (*P = 0.0123) [two-tailed unpaired t test with Welch’s correction: mean ± SEM, n = 15/condition, 3 clutches]. A, B, E, G, H–J, L, M Source data are provided as a Source Data file.