Fig. 2

Defective Kmt2a expression decreased the proliferation of neural progenitors. (A) In situ hybridization of 75%-epiboly embryos showed that sox2 expression was downregulated by the kmt2a morpholino or kmt2a^N84 cRNA, as confirmed by qPCR analysis on the right. (B) Embryos were flat-mounted and double-labeled with sox2 and phospho-histone H3 antibody. The bottom panels are representative of the enlargement regions of the upper panels, as indicated. sox2-expressing cells were pseudo-colored with fluorescent red and counterstained with phospho-histone H3 antibody for immunohistochemistry (fluorescent green) to locate proliferating neural progenitor cells (arrowheads). Injection of the kmt2a morpholino or kmt2a^N84 decreased proliferation of neural progenitors. The proportions of phospho-histone H3- and sox2-positive cells among the total sox2-positive cells were quantified, as shown on the right. Note that in the Kmt2a-deficient embryos, no significant deviation was observed in the proportions of phosphohistone H3-positive and sox2-negative cells in the sox2-negative cells counted in adjacent surface ectoderm.