Figure 6.

Immunofluorescence staining of P5 Dicer CKO + NICD retinas. A–F, nuclear GFP staining (green) indicates areas of Dicer CKO and NICD expression (CKO+ NICD). Note that scattered GFP⁺ cells are present in wild type areas of retina due to αPax6 expression in a subset of amacrine cells. A–E, left panels focus on wild type expression of indicated markers. A′–E′, right panels focus on CKO+NICD expression of indicated markers. A–A′, EdU was injected at P3. CKO+NICD areas contain cells colabeled with the ganglion cell marker Brn3 (red) and EdU (blue), indicating birth at P3. B–B′, CKO+NICD areas contain cells that express normal levels of Sox2 (red), and show progenitor morphology. C–C′, CKO+NICD areas contain progenitor cells that express Id1 (red) at lower levels that nearby wild type Müller glia. D–D′, CKO+NICD areas lack expression of the Müller glial marker Cralbp (red). E–E′, Costaining with Prox1 (red) and neurofilament (NF, blue) reveals the presence of horizontal cells in CKO+NICD areas, though these cells are absent from Dicer CKO areas at P5 (Georgi and Reh, 2010). F, Occasional groups of CKO+NICD cells show induction of gliogenesis (arrow) as indicated by Cralbp staining (red). These cells are almost always located in close proximity to wild type cells. Scale bars: 100 μm. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]