Figure 5.

Melanoma exosomes modulate endothelial cytokine production. 2F-2B endothelial cells (50,000/well) were cultured for 72 hrs to form spheroids followed by the addition of 0, 2.5, or 10 μg/ml for 10 days to produce cytokines in the spheroid culture media. Three sets of culture media samples were collected and pooled for each exosome dosing group and analyzed using cytokine arrays. (a) TransSignal Mouse Angiogenesis Antibody Array (Panomics) or (b, c) Proteome Profiler Mouse Cytokine Array (R&D Systems). Cytokines levels are represented as a percent change over control (control = 0). The absolute change in level for each cytokine group (2.5 and 10 μg/ml melanoma exosomes) was averaged. The average of absolute changes for all cytokines was 41%. ANOVA analysis was used to calculate statistical significance. Cytokine levels that changed more than 10% from the absolute value average changes as compared to a normalized control group were considered to be affected by the experimental procedures, according to ANOVA that revealed statistical significance (p < 0.05) for difference from control in each case. The p values for 2.5, 10 and combined are 0.035, 0.016 and 0.001 respectively. Error bars = s.e.m. of replicates present on the arrays.