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. Author manuscript; available in PMC: 2014 Jan 1.
Published in final edited form as: Nat Cell Biol. 2013 Jun 2;15(7):807–817. doi: 10.1038/ncb2767

Figure 5.

Figure 5

Notch1-mediated reduction in neovascular tips suppresses breast tumor cell outgrowth. Microvascular niches were created with stromal cells mixed with shCtrl E4-EC and/or shNotch1 E4-EC. YFP T4-2 cells were then seeded in SFM and growth was analyzed 10 days later. (a) Microvascular niches composed of shCtrl E4-EC, shNotch1 E4-EC, or a 1:1 mix of the two (‘sh1:1’) were fixed and stained for CD31 at day 7. Scale bar = 200 μm. (b) Neovascular tip number/field (large white dots in a) and (c) branch point density (small yellow dots in a) were quantified (n=15 fields of microvascular networks pooled from 5 separate co-cultures). Error bars denote s.e.m ***p<0.001 when compared to shCtrl condition by one-way ANOVA and Dunnett’s post-test. (d) Representative images of YFP T4-2 seeded upon shCtrl EC, shNotch1 EC, or sh1:1 cultures, fixed and stained for CD31 and Ki67 10d post-seeding. Scale bar = 200 μm. (e) Quantification of normalized tumor cell area fraction and (f) %Ki67-negative clusters (each normalized by shCtrl condition; n=5 sets of co-cultures analyzed per condition). Error bars denote s.e.m. *p<0.05 when compared to shCtrl condition by one-way ANOVA and Dunnett’s post-test in (e).