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. 2021 Mar 5;22(5):2607. doi: 10.3390/ijms22052607

Figure 6.

Figure 6

Retinal differentiation potential following NR2E3 allele knockout. Bright-field images of WT (A), G56R (E), and G56R-CRISPR2 (I) iPSC-derived retinal organoids at 180 days of differentiation. IF analysis of NR2E3 expression (in green) in WT (B), G56R (F), and G56R-CRISPR2 (J) retinal organoids; nuclei labelled in blue. NR2E3 expression is localized to the photoreceptor nuclei, which form an outer layer, corresponding to the outer nuclear layer (ONL), in the organoid. Colocalization studies of NR2E3 (in green) and OTX2 (in red) expression in WT (C), G56R (G) and G56R-CRISPR2 (K) retinal organoids. NR2E3 is restricted to the rod nuclei whereas OTX2 also labels the more peripheral cone nuclei, as well as retinal cells situated on the inner side of the ONL. Colocalization studies of CRX (in green) and NRL (in red) expression in WT (D), G56R (H), and G56R-CRISPR2 (L) retinal organoids. NRL expression is restricted to rod nuclei, whereas CRX also labels the cone nuclei situated on the outer rim of the organoids.