Retina-like tissue in the subretinal space of a 2-week-old PRPE65-ngn3 mouse.

A–C: Low magnification images to show the relative position of retina-like tissue (arrow) in a cross-section of the eye, with the anterior to the left and the posterior to the right. A: Bright-field (BF) view. B: Epi-fluorescence of DAPI staining of the nuclei. C: Epi-fluorescence of anti-recoverin (Rcv) immunostaining to mark photoreceptor cells. Boxes outline the corresponding regions in A–C, or similar locations on serial sections (D–O). D–G: A serial section under bright-field view (D), after DAPI nuclear staining (E), and after double immunostaining for Otx2 (in green, F) to identify bipolar cells and for Pax6 (in red, G) to label amacrine cells in the INL and in the GCL (misplaced amacrine). H–K: A serial section under bright-field view (H), with DAPI nuclear staining (I), and with double immunostaining for AP2α (in green, J) to label horizontal (albeit weakly) and amacrine cells in the INL and in the GCL (misplaced amacrine), and for Brn3A (in red, K) to identify a subset of ganglion cells. L–O: A serial section under bright-field view (L), with DAPI nuclear staining (M), and with double immunostaining for photoreceptor protein recoverin (in green, N) and for RPE protein RPE65 (in red, O). RPE-L, RPE-like; ONL-L, outer nuclear layer-like; INL-L inner nuclear layer-like; GCL-L, ganglion cell layer-like. Scale bar (100 μm) in A also applies to B, C. Scale bar (100 μm) in D also applies to E–O.