|
The maturation of erythrocytes is disturbed in prpf4 mutants. A, B Morphological analysis of erythrocytes in siblings and prpf4−/− embryos at 54 hpf. A Representative confocal images showing GATA1+ cells stained with DAPI in siblings and prpf4−/− embryos. In sibling embryos, 81.7% of erythrocytes exhibit an oval morphology, while in prpf4−/− embryos, 53.1% of erythrocytes display a round morphology. The presented images represent typical erythrocyte morphologies observed in each group. Scale bar: 20 µm. B Quantification of the nuclear-to-cytoplasmic (N/C) ratio of individual GATA1+ cells (30 embryos per group). C WISH analysis of gata1a expression in siblings and prpf4−/− embryos at 36 hpf and 40 hpf. Lateral views (c3, c4) and ventral views (c3’, c4’). Scale bar: 200 µm. D Relative expression of gata1a in prpf4−/− mutants and siblings at 36 hpf and 40 hpf, as determined by RT-qPCR. E RT-qPCR analysis of erythroid-related gene expression in siblings and prpf4−/− embryos. F WISH analysis of hbbe3 expression in siblings and prpf4−/− embryos at 54 hpf. G Combined analysis of hbbe3 expression and hemoglobin levels in embryos. g1, g2 O-dianisidine staining reveals reduced hemoglobin levels in prpf4−/− embryos compared to siblings. g1′, g2′ Whole-mount in situ hybridization for hbbe3 performed on the same embryos as in (g1) and (g2). Scale bars: 100 µm (F,G).
|
