Supplementary Figure S3 from Targeting Dendritic Cell Dysfunction to Circumvent Anti-PD1 Resistance in Head and Neck Cancer

Figure S3 A. Representative flow cytometry results of isolated Xcr1+ cDC1s used for DC vaccine experiments. B. Histogram of indicated marker expression gated on Xcr1+ cDC1s analyzed by flow cytometry. Xcr1+ cDCs were isolated and cultured with (=activated) or without (=no stimulation) PolyI:C (20 μg/ml) for 4 hours before staining. C, D. IFN-γ ELISA testing the ability of Xcr1+ DCs to activate CD8+ T cells. CD8+ cells isolated from OT-1 mouse splenocyte (OT1) were co-cultured with Xcr1+ cDC1s (DC), with or without stimulation of ovalbumin short peptide (short), ovalbumin synthetic long peptide (SLP) in Figure S3C and MOC1esc1-OVA (full length) cell lysate (MOC1esc1OVA) in Figure S3D. n=2-3. E, F. Flow cytometric analysis of MOC1esc1 tumors (E) and DLNs (F) treated with intra-tumoral PBS or DC vaccine on days 1/4/7 post inoculation, and harvested on day 14 post tumor inoculation. (n=8, representative data of two independent experiments.) G. Tumor weight measured on last day of experiment in Figure 3E. H. Photo of tumors harvested in experiment shown in Figure 3E