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Radial Glial Neural Progenitors Regulate Nascent Brain Vascular Network Stabilization Via Inhibition of Wnt Signaling

Figure 5

Increased proliferation and up-regulated canonical Wnt pathway activity in ECs following neural progenitor ablation.

(A–B′) Cortical EC proliferation at E16.5. BrdU labeling (in red) revealed increased clusters (arrowheads) of BrdU+ ECs (IB4 in green) in mutants (B and B′). (C–D′) Ki67 staining at E16.5. Increased clusters of Ki67+ ECs (in red, arrowheads) were observed in mutants (D and D′). (E–F) Glut-1 expression in ECs at E16.5. Up-regulation of Glut-1 expression (in red) was observed in mutants (F). (G–H) Expression of Wnt reporter BAT-lacZ in ECs at E16.5. X-Gal reaction revealed increased numbers of lacZ+ (in blue, arrowheads) ECs (IB4 in brown) in mutants (H). BAT-lacZ up-regulation in neural cells appears largely restricted to the intermediate zone (IZ). (I–J) Western blot analysis of Glut-1 expression at E16.5. Stronger bands were observed in mutants (I). Quantification showed a >150% increase in mutants (J) (p = 0.03, n = 3). (K–L) Quantification of EC proliferation and BAT-lacZ expression at E16.5. The density of both BrdU+ (p = 0.001; n = 4) and lacZ+ (p = 0.001; n = 3) ECs is significantly increased in mutants. Scale bar (in D): 200 µm for (A–H).

Figure 5