Apoptotic potential of TRAIL-producing ADSCs and the effect of IFNγ on sensitivity of melanoma B16F10 cells to TRAIL in co-culture experiments. A) WT-ADSCs and TRAIL-ADSCs co-cultured with CAOV-4, Ej-138, MCF-7 and B16F10. Upper panel: Representative microscopy images from each one of triplicate co-culture experiments. Inlays show the morphology of each cancer cell line. Lower panel: Representative FACS plots showing a statistically significant increase in dead CAOV-4 and Ej-138 cells (Student t test, P < .001), implying bioactivity of TRAIL produced by murine modified ADSCs. Each plot represents a mean value of three independent experiments. P values are two-sided. Scale bars =50 μm. B) FACS plots from co-culture experiments demonstrated a decrease in proliferation of red fluorescent protein (RFP)-expressing melanoma cells in groups co-cultured with WT-ADSCs and TRAIL-ADSCs only in the presence of IFNγ. Population of ADSCs and melanoma cells are shown in black and red respectively. One experimental representative of triplicate independent experiments is shown. C) A FACS analysis (results depicted as bar graph) demonstrated a statistically significant decrease in melanoma cell number co-cultured with both WT-ADSCs and TRAIL-ADSCs after addition of IFNγ-containing CM. However, the effect of IFNγ at 48 h of incubation on melanoma cells co-cultured with WT-ADSCs was more significant (compared to melanoma cells co-cultured with TRAIL-ADSCs), implying IFNγ does not effect the activity of TRAIL on melanoma B16F10 cells. Experiment was performed in triplicate. Error bars correspond to 95% confidence intervals. P values are two-sided. ***P < .001 (ANOVA). WT = wild type.