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Fig. 7. Shh maintains the RPE phenotype and its actions in the RPE and the CB/CMZ depend on Fgfr signaling. (A) Developing eyes were treated with Fgf2 at E4, or Rcas-Shh and Fgf2 at E3.5 and E4, respectively, and analyzed at E6. Ectopic Shh was sufficient to inhibit transdifferentiation in developing eyes. B, Fgf2 bead; t, transdifferentiated retina. Arrowhead indicates developing retina. (B) Ffgr signaling activity was measured 4 hours after treatment by immunohistochemistry using a phospho-ERK antibody in retinectomized eyes with: no Fgf2, Fgf2, Fgf2/KAAD or Fgf2/Rcas-Shh. Fgf2 activates the Fgfr signaling pathway, whereas ectopic Shh inhibits this activity. Scale bar: 50 µm. (C) Retinectomies were performed at E4 and eyes were treated with Fgf2±PD 173074, or with Rcas-Shh±PD 173074, and analyzed 3 da ys later. Ectopic Shh stimulates regeneration from the CB/CMZ, as does Fgf2. Both of these activities are significantly inhibited when PD 173074, a potent and specific Fgfr inhibitor, is used.





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