Fig. 4

Gynohaploid development of asf1 KD embryos. a A group of syncytial nuclei in metaphase from an asf1 KD embryo. These nuclei are haploid (four chromosomes), as visible in the close-up. Scale bar: 10 μm. b Confocal sections of cellularized asf1 KD (n = 67) and control blastoderm (n = 83) embryos. Insets were taken at the surface of the same embryo (top) or in the same plane than the main image (bottom). asf1 KD embryos typically show irregular organization of the nuclear layer, with frequent patches of nuclear sinking (yellow arrows). Note the doubled nuclear density at the surface in haploids compared to the diploid control. Scale bar: 50 μm. c In control eggs (left) at pronuclear apposition, ASF1 (green) is abundant in both apposed pronuclei (100%; n = 20). In asf1 KD eggs (right) at the same stage, no signal is detected in apposed pronuclei (0%; n = 24). Note that the female pronucleus appears normal while the male pronucleus fails to decondense (arrowhead). Scale bar: 10 μm. Color panels in c show merged stainings