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Fig. 5 Definitive granulopoiesis and thrombopoiesis are boosted in Trim33-deficient mutants. (A) Lateral view of the trunk (left panel) and CHT (right panel) at 5 dpf of control siblings (top) and monNQ039 mutants (bottom) expressing the coro1a:eGFP (leucocytes, green channel) and lyz:DsRed2 (neutrophils, red channel) transgenes. (B) Quantification of neutrophil-associated fluorescence in live monTB222/Tg(mpx:eGFP) siblings and mutants from 5 to 9 dpf, based on total fluorescence (sum of pixel intensities per larva); n=33 mutant and 33 wild-type larvae. Data are mean±s.e.m. (C) Whole-mount Sudan Black staining of mature neutrophils in monNQ039 siblings (top) and mutants (bottom) at 7.5 dpf; lateral view of the kidney area – rostral towards the left. (D,E) Lateral view of the cardiac (D) and caudal (E) regions of 6 dpf live control sibling (left) and mutant (right) larvae expressing the gata1:DsRed (erythrocytes and thrombocytes, red channel) and CD41:eGFP (HSPCs and thrombocytes, green channel) transgenes, pre-treated with BDM to stop the heartbeat prior to imaging. (F) Caudal region of 3 dpf zebrafish Tg(CD41:eGFP) larvae control sibling and mutant larvae. (G) Kidney region of 5 dpf Tg(CD41:eGFP) larvae control sibling and mutant larvae. Sib, sibling; Mut, mutant; k, kidney; sb, swim bladder; h, heart; da, dorsal aorta; av, axial vein; ca, caudal artery; CHT, caudal hematopoietic tissue; cv, caudal vein; nc, notochord; g, gut. Scale bars: 50 µm.