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. 2017 Jan 28;25(3):634–653. doi: 10.1016/j.ymthe.2016.12.015

Figure 2.

Figure 2

Investigation of the Chrnb4-EGFP-Positive Cell Proliferation during the Cone Generation Wave

Pregnant females were labeled for 24 hr with EdU at E12, E13, E15, or E17. (A–DII) Transversal sections are shown of retinas at E13, E14, E16, and E18 stained for EGPF and EdU. (A–D) Merged pictures show EGFP (green) and DAPI nuclear staining (in blue). (AI–DI) EdU labelings (red) at different developmental stages are shown. (AII–DII) Merged pictures show EdU-positive cells (red) and EGFP-positive cells (in green). Dashed lines delimit the apical (Ap) from the basal (Ba) retinal regions used for counting in (F) and (G). (E–G) Percentage of double EdU-/EGFP -positive cells were counted over the EGFP-positive cells detected in the whole (E), apical (F), and basal (G) retina. (H–J) Percentage of double Ki67- and EGFP-positive cells counted over the total number of EGFP-positive cells confirms the results achieved with the EdU labeling. DAPI, nuclear staining (blue); Ba, basal; Ap, apical. (E–G and J) ANOVA with Tukey’s correction, ***p < 0.001; n = 4. Error bars correspond to SEM. Scale bars, 25 μm (A–D, H, AI–DI, and HI) and 15 μm (AII–DII and HII).