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Figure 5


Fig. 5. Identification of transdifferentiated precursor cells in Ptf1a-deficient retina at E18.5. (A-F) Sections from E18.5 retinae of Ptf1a+/Cre(ex1) Z/EG and Ptf1aCre(ex1)/Cre(ex1) Z/EG were double-immunostained with anti-GFP and anti-Brn3 antibodies. (A,D) Ptf1a-mediated GFP expression is present in Ptf1a+/Cre(ex1) Z/EG and Ptf1aCre(ex1)/Cre(ex1) Z/EG retinae (green). (B,E) Ganglion cells are marked with Brn3 (red). (C,F) Yellow in merged images represents co-localization of GFP with Brn3. GFP coexpression with Brn3 is not present in Ptf1a+/Cre(ex1) Z/EG retina (C). By contrast, in about one-third of Ptf1aCre(ex1)/Cre(ex1) Z/EG retinal cells, GFP is coexpressed with Brn3 (F, arrows, inset). (G,H) Quantification of Brn3-positive cells in wild-type (wt) and Ptf1aCre(ex1)/Cre(ex1) (Cre/Cre) retinae (G) and of GFP and GFP/Brn3-positive cells in Ptf1aCre(ex1)/Cre(ex1) Z/EG retinae (H). Each histogram represents the mean±SD for four retinae (Number of Brn3-positive cells in wt are 28.5±2.08 and in Cre/Cre are 40.75±3.40. In Ptf1aCre(ex1)/Cre(ex1) Z/EG retinae, the number of GFP-positive cells are 27.2±1.70 and GFP/Brn3-positive cells are 9.2±3.59). Positive cells were scored in an area that was above a line of 200 µm in length parallel to the retinal pigment epithelium. Scale bars: 50 µm. GCL, ganglion cell layer; NBL, neuroblastic layer.