Figure 3.

Effects of CAFs on DC activation markers. Immature DC (iDCs) stimulated with a maturation cytokine cocktail were incubated for 48h with conditioned medium from irradiated or non-irradiated CAFs (CAF-CM, left panels) or in co-cultures (CAF-CC, right panels). Resulting expression of mDC cell surface markers CD40, CD80, CD86, and HLA-DR was evaluated by flow cytometry. (A) Gating strategy used to analyze the expression of activation markers in DCs. (B) Representative histograms of expression by mean fluorescence intensity (MFI) of CD40, CD80, CD86, and HLA-DR in DCs stimulated with maturation cocktail in culture with conditioned medium from non-irradiated and irradiated CAFs (1 donor). (C) Bar graphs represent mean ( ± SD) values from flow cytometry analysis of four-4 different CAF donors, measured independently. Pattern columns indicate protein surface levels in control iDC and mDC cultures. Results are expressed as percentage of total cells. Data represent mean ( ± SD) values from 4 different CAF donors measured independently. Brown-Forsythe and Welch ANOVA test and p-values were determined between control and non-irradiated CAFs, mDCs, and the two irradiated CAF-groups individually. *p ≤ 0.05, **p ≤ 0.01.