Figure 8

In vivo analysis of MNTX synergy with temsirolimus on inhibition of angiogenesis. In Panels A and B, 10-week-old female C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME) were injected subcutaneously on the ventral abdomen with 500 μl Matrigel (BD Biosciences, San Jose, CA) containing either MNTX (100 nM), temsirolimus (10 nM), or both drugs (100 nM MNTX and 10 nM temsirolimus). 20 ng VEGF was added to all Matrigel plugs. After 21 days, the plugs were removed. Panel A: Pictures of Matrigel plugs after removal from mice. Panel B: Graphical analysis of Matrigel plug hemoglobin content. The plugs were weighed and homogenized, and their hemoglobin content was quantified using the QuantiChrom™ hemoglobin assay kit (BioAssay Systems, Hayward, CA), n = 3 per group. The asterisks indicate a statistically significant difference (p < 0.05) between treatment and control. There is also a statistically significant difference (p < 0.05) between MNTX or temsirolimus treatment alone versus in combination.