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. Author manuscript; available in PMC: 2012 Mar 30.
Published in final edited form as: Lab Invest. 2009 Sep 28;89(11):1317–1328. doi: 10.1038/labinvest.2009.94

Figure 4.

Figure 4

Long Term Effects of varying B16-F10 melanoma exosome concentrations on 3D endothelial culture. 2F-2B endothelial cells (50,000/well) were cultured for 72 hrs to form spheroids followed by the addition of 0, 2.5, 5, or 10 μg/ml for 10 days to study the permanence of exosome effects on spheroid development. (a) phase contrast microscopy of representative spheroid morphology for each exosome dose. (b) ImageJ software was used to measure the area of individual spheroids in 40 X mag. fields (see supplemental information, Fig. S1). Average spheroid size/field was normalized against control (0 μg). ANOVA was used to calculate significance for p < 0.05 for a control of N = 48; * = p = 5.4×10^−5. Error bars represent the standard deviation of the average spheroid size from 3 random fields from different cultures. (c) XTT (tetrazolium salt) colorimetric reagent (MD Biosciences; St. Paul MN) was used to measure cell proliferation. Average endothelial cell proliferation was normalized against control (0 μg exosomes). ANOVA was used to calculate significance for p < 0.05 for a control of N = 4; ** = p = 5.9×10^−3 and *** = p = 2.9×10^−3. Error bars represent the standard deviation of N = 4 samples.