Figure 6

Correlation of Pax6 changes in G2/M phase with the emergence of a neuronal marker. (A-C) Confocal images of Pax6 and PH3 co-labeling at E6.5 (HH stage 30). Boxed region in (A) is shown as Pax6 and PH3 merge (B) or Pax6 alone (C) at twofold magnification, where dotted outlines in (C) encircle all PH3⁺ cells. (D) Flow cytometry profiles of Pax6, PH3, and DNA content at E7 (HH stage 32). For both panels the y-axes represent Pax6 levels, while the x-axes indicate the DNA content. The left panel shows merged distribution profiles of Pax6 (red) and PH3 (green), and the percentage of PH3⁺ cells among total cells is indicated below. The right panel shows cell density distribution with regard to Pax6 and the cell cycle. The percentages of Pax6^Hi, Pax6^Lo, and Pax6^Neg cells among total cells are indicated at the right. (E-H) Confocal images of Visinin, PH3, and DAPI co-staining at E6.5 (HH stage 30). The boxed region in (E) is shown as Visinin- (F), PH3- (G), or DAPI staining (H) alone at twofold magnification. (I-L) Co-labeling for Visinin (J), PH3 (K), DAPI (L), and merge (I) in dissociated E6 retina cells. (M-P) Confocal images of Visinin, Pax6, and DAPI co-staining at E6.5 (HH stage 30). The boxed region in (M) is shown as Visinin (N), Pax6 (O), or DAPI staining (P) alone at twofold magnification. Yellow dotted lines encircle Visinin⁺Pax6^Neg cells, whereas white dotted lines in (N-P) encircle a pair of cells at the end of the M-phase with weak Pax6 and Visinin signals. Arrows indicate co-labeled cells. Arrowheads point to marker-positive but Pax6^- cells. vs, ventricular surface. Scale bars: 100 μm.