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Fig. 5. Manipulation of Shh levels. (A) Ectopic expression of Shh using Rcas-Shh. (A1) Rcas-Shh/Fgf2-treated eyes show an increase in the expression of Shh mRNA (arrowhead). (A2) Shh sense probe. (A3) Ectopic Shh expression upregulates Ptc1 levels as indicated by the arrowhead. (A4) Ptc1 sense probe. (A5) Ectopic Shh expression via a retroviral vector corresponds to an increase in Shh protein (arrowheads). (A6) AMV3C2 staining shows the presence of Rcas virus coat protein in Rcas-Shh-treated eyes (arrowheads). (B) Real-time PCR analysis of the relative Ptc1 levels in Fgf2, RCAS-Shh/Fgf2, and 100 µM KAAD/Fgf2-treated eyes show that Rcas-Shh treatment causes and significant increase in Ptc1 (P<0.01) levels while KAAD/Fgf2 treatment reduces Ptc1 levels (P<0.05). * statistically significant. Error bars are s.e.m.





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