Fig 2: The effect of PBNP-PTT-administered thermal dose on co-stimulatory and HLA molecules is more pronounced in SH-SY5Y cells compared with LAN-1 cells. (A–D) SH-SY5Y and (E–H) LAN-1 cells were subjected to various thermal doses via PBNP-PTT. After treatment, cells were rested for 24 h and evaluated by flow cytometry analysis for (A,E) CD80, (B,F) CD86, (C,G) HLA-ABC, and (D,H) HLA-DR. MFI is represented as bar graphs representing averages, and corresponding representative histograms. Inset values in the histograms denote the thermal dose. Each parameter was compared to the vehicle control by one-way ANOVA, where * p < 0.03, ** p < 0.002, *** p < 0.0002, **** p < 0.0001; n = 2/group.

Fig 3: Single cell NGS analysis of CD68/CD163 MΦ clusters in 2 datasets (GSE139324 and GSE164690) of human HNSCC. (A) Quality control (QC), metrics and selection of CD14+|CD11b+, MPO-|CD123-|CD1C-, APOE+|MRC1+ cells before further analysis. (B, C) UMAP visualization of the CD68/CD163 MΦ clusters of human HNSCC (study A (GSE139324): 3446 cells, study B (GSE164690): 3930 cells). (D, E) Violin plots of the expression levels of CD14, CD11c, CD33, CD68, CD163, PD-L1, CD64, CD80 and CD206 within the clusters of A (GSE139324) and B (GSE164690). (F, G) Color-coded feature plot of CD68 and CD163 expression per cell projected onto the UMAP visualization of A (GSE139324) and B (GSE164690).

Fig 4: Flow cytometric characterization of CD68/CD163 subsets in HNSCC. (A) Schematic overview. Single cell suspensions of 3 primary human HNSCCs were labeled using fluorochrome-conjugated antibodies. Cell differentiation was performed according as shown. The detailed gating can be found in Supplementary Figure 5 . (B) Exemplary dot plot of the distribution of leukocyte populations within CD68/CD163 subsets. (C) Proportion of leukocyte populations within the CD68/CD163 subsets. B Exemplary dot plot of the distribution of leukocyte populations within CD68/CD163 subsets. C Proportion of leukocyte populations within the CD68/CD163 clusters. (D) Proportion of PD-L1-expressing leukocytes to total PD-L1 expression. (E) Proportion of PD-L1 expressing cells within the CD68/CD163 subsets. (F) Proportion of PD-L1-expressing leukocyte subtypes to PD-L1 expression of the respective CD68/CD163 subset. (G) Exemplary dot plot of CD80 and CD206 expression of leukocyte populations. (H) Proportion of CD80 expressing cells within the CD68/CD163 subsets. (I) Proportion of CD80-expressing leukocytes to total CD80 expression within the CD68/CD163 subsets. (J) Proportion of CD206 expressing cells within the CD68/CD163 subsets. (K) Proportion of CD206-expressing leukocyte subtypes to CD206 expression of the respective CD68/CD163 subset. Within bar plots each bar represents the mean value of three samples. The error bars show the standard error.