Fig. 6: Functional characterization of cDC2s and CD14⁺ DCs isolated from human OMC.

a Two days after total cDC2s injection, BLM-OMCs were digested and CD1c⁺CD14⁻ (cDC2s) and CD1c⁺CD14⁺ (CD14⁺DCs) subsets FAC-sorted for RNA extraction and molecular characterization by qRT-PCR. Gene expression levels (2^−ΔCt) for the indicated genes in cDC2s and CD14⁺DCs. Reported values for each gene are the means of n = 3 biological replicates (mean ± SEM; two-way ANOVA and Sidak’s multiple comparisons test). ACTB was used as internal reference. Colour legends indicate: cDC2s (orange), CD14⁺DCs (green). b IL-6 production assayed by intracellular cytokine staining in BLM-educated cDC2s, upon stimulation with LPS (1 μg/mL) for 6 h. cDC2s and CD14⁺DCs were identified and gated based on CD1c and CD14 expression in CD45⁺ live single cells. Representative dot plots are shown. Numbers indicate the percentage of gated cells. c Intracellular protein expression of S100A9 in tumour-educated cDC2s and CD14⁺DCs. Representative GeoMFI histograms for BLM and Mel624 are shown. d Proliferation of allogeneic CD3⁺ T cells 5 days after co-culture with FAC-sorted cDC2s and CD14⁺DCs. Scattered dot plots (five independent experiments, in triplicate (Exps. 1–2-3) and duplicate (Exps. 4-5) samples; mean ± SEM; one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons tests). Representative CFSE histogram plots from Exp. 2 are shown; numbers indicate the percentage of gated cells. e Biplot of HLA-DR expression and induced allogeneic T-cell proliferation in Exps. 4 and 5. Correlation assessed by linear regression. f Autologous T cells were co-cultured with cDC2s and CD14⁺DCs for 5 days and then stained for HLA-DR, CD25, CD3, CD8 and live/dead marker. Percentage of activated CD25⁺HLA-DR⁺ cells in total CD3⁺ T cells is reported. Scattered dot plots (two independent experiments, in triplicate samples (mean ± SEM; one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons tests). Representative dot plots from Exp. 1 are shown. Statistical significance was annotated as follows: **p < 0.01, ***p < 0.001, ****p < 0.0001. Source data are provided as a Source data file.