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. 2009 Aug 25;106(39):16698–16703. doi: 10.1073/pnas.0905245106

Fig. 3.

Fig. 3.

Acquisition of optic vesicle and optic cup cell phenotypes. (A) Mitf protein expression in neurospheres after 30 days of differentiation. (B) qPCR analysis of Mitf gene expression over the first 80 days of differentiation. (C–E) Immunocytochemical analyses of the time course of Mitf and Chx10 protein expression in neurospheres at 30 (C), 40 (D), or 50 (E) days of differentiation. (F) qPCR analysis of Chx10 gene expression over the first 80 days of differentiation. (G) Uniform Chx10 expression throughout a subset of neurospheres by day 40. (H) FACS analysis demonstrating the percentage of all cells expressing Chx10 at day 40. (I) Immunocytochemical analysis showed all Chx10+ cells coexpressed Pax6 at day 40. (J) Rosettes of Chx10+ cells expressed the tight junction protein ZO-1 within their core. (K) Rare Chx10+ cells coexpressed βIII tubulin at day 40. (L) qPCR demonstrating increased Mitf expression and corresponding decreased Chx10 expression in adherent cultures treated with the FGF inhibitor SU5402. qPCR values were expressed as fold change relative to cultures at day 16 (B and F) or day 10 (L) of differentiation. (Scale bars, 500 μm in panels A and G; 50 μm in panels C–E, J, and K; and 75 μm in panel I; blue stain in A and G is Hoechst nuclear dye.)