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Figure 6


Fig. 6. Angiopoietin signaling regulates Wnt7b expression and cell cycle stage-dependent cell death. (A,B) X-Gal staining of hyaloid vessels from Ang2lacZ/+; Wnt7blacZ/+ (A) and Ang2lacZ/lacZ; Wnt7blacZ/+ (B) animals. Macrophages are circled in red. (C) Wnt7b and Gapdh RT-PCR of laser captured macrophages from hyaloid vessels of P3 wild-type animals, injected intra-vitreally with Ang2 (Ang2) or sham injected (con). Data from three independent experiments are presented. (D) Ang1 and Gapdh RT-PCR from wild-type P5 hyaloid vessel mRNA. Whole eye RNA was used as a control. (E-H) Combined BrdU (green) and TUNEL (red) labeling of hyaloid vessels showing BrdU-positive VECs that are apoptotic (F, indicated by white arrowhead) and pairs of BrdU-positive daughter nuclei (G,H). (I,J) BrdU labeled cell fate mapping: red lines show the number of BrdU-TUNEL double labeled nuclei (I) and BrdU-positive daughter pairs (J) in left eye hyaloid vessels assessed over a 21 hour time-course after BrdU incorporation. The blue lines in I and J show similar quantification in the right, contralateral eye after Ang1 injection at t=6 hours. Significance levels: **P≤0.01, >0.001; ***P≤0.001, >0.





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