Figure 1.

Spatiotemporal Distribution of K14⁺ Precursor Cells during Embryonic and Neonatal Development

(A–C) Confetti embryos (n=4 eyes/time point) harvested at E16.5 (A), E18.5 (B), and E19.5 (C) with corneas imaged by confocal microscopy. Optical cross-sections (A–C, lower panels) were acquired at the plane marked by thick hatched lines. Thin circular hatched line represents the cornea boundary.

(D–F) Neonatal corneas (n = 4 eyes/time point) imaged at P7 (D) and P14 (E and F). An optical cross-section (D, lower panel) was obtained along the plane marked by the thick hatched line. Whole corneas from P14 mice were flat-mounted (E and F). Solid white lines (E) depict incisions made to facilitate flat-mounting. The hatched box (E) is magnified in (F). Optical cross-section (F, lower panel) was obtained at the plane marked by the solid white line.

(G) Total number of fluorescent cells within developing mouse corneas (n = 4 corneas/time point, ± SD, ^∗∗p < 0.01 and ^∗∗∗p < 0.001, one-way ANOVA with Holm-Sidak multiple comparison test).

(H) Clonal size represented by the number of cells (n = 10 clones/cornea, ± SD, ^∗p < 0.05 and ^∗∗∗∗p < 0.0001, one-way ANOVA with Holm-Sidak multiple comparison test).

(I) Representative cryo-section prepared from P14 mouse eye; only RFP (red) is displayed, and tissue was counterstained with DAPI (blue) to highlight cell nuclei. The white hatched line represents the epithelial basement membrane.

Scale bars: (A–D) 300 μm, (E) 400 μm, (F) 30 μm, (I) 10 μm.