Fig. 5

IFNG neutralization impacts T cell effect on MDMs. (A) Primary M0 MDMs were cultured stepwise in mono-culture, to co-culture with established LNCaP or 22Rv1 tumor cultures; T cells were added for the last 24 h with vehicle control (Ctrl), or IgG control (IgG Iso), or anti-IFNG neutralizing antibody (anti-IFNG). (B) IFNG protein concentration (pg/ml) in cell culture supernatants; n = 5; data expressed as mean ± SEM. (C) MDM mRNA expression in the six tri-culture cellular and treatment conditions shown as normalized relative quantity (NRQ) as related to house-keeping genes RPLP0 and POLR2A. One-way ANOVA nonparametric with Friedman post-hoc analysis comparing the mean rank of each column with the rank of every other column; n = 5; Data represent mean ± SEM. (D,F) Concentrations of secreted cytokines and chemokines in supernatant from the six tri-culture cellular and treatment conditions, expressed as pg/mL. One-way ANOVA nonparametric with Friedman post-hoc analysis comparing the mean rank of each column with the rank of every other column; n = 5; Data represent mean ± SEM. (E) T cell migration analysis measured by in-chip confocal microscopy imaging in individual microdevice wells. Data is expressed as each sample’s fold change in migration distance as relative to its corresponding tri-culture vehicle control condition quantified by NIS-Elements. One-way ANOVA nonparametric with Friedman post-hoc analysis comparing the mean ranks of vehicle control IgG control, and vehicle control to Anti-IFNG neutralizing antibody; n = 5; Data represent mean ± SEM.