Additional file 1 of Myeloid cell-derived apCAFs promote HNSCC progression by regulating proportion of CD4+ and CD8+ T cells
Creators: name: Ren, Feilong
name: Meng, Lin
name: Zheng, Shize
name: Cui, Jiasen
name: Song, Shaoyi
name: Li, Xing
name: Wang, Dandan
name: Liu, Qilin
name: Bu, Wenhuan
name: Sun, Hongchen
Publisher: name: figshare
Year: 2025
Subjects: subject: Biochemistry
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Pharmacology
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Hematology

description: Supplementary Material 1: Fig. 1. HE staining of tumor tissues from different HNSCC models in both patients and mice. A, Hematoxylin and eosin (H&amp;E) stained histological sections of tumor tissues from patients with OSCC used in this study. B, H&amp;E staining histological sections of OSCC samples induced by 4NQO in mice. C, H&amp;E staining histological sections of murine transplant tumors following inoculation with the mouse OSCC cell line SCC7. Fig. 2. Successfully established a myeloid cell-specific fluorescent reporter mouse and an OSCC model. A, Tissue specimens of normal mouse tongue and 4NQO-induced OSCC in mouse tongue. B, Genotyping results for Lyz2-Cre/ROSA-mTmG mice. C, Fluorescence identification results of spleen in Lyz2-Cre;ROSA-mTmG mice. Fig. 3. Single-cell data and TCGA database analysis results. A, the expression levels of different fibroblast-associated markers (COL3A1, COL1A1, PDGFRB, FAP) and MHC class II-related markers (HLA-DPB1, HLA-DRA, HLA-DPA1, CD74) across three CAFs subpopulations in HNSCC. B, Pseudotime analysis results of epithelial cells and CAFs in human and mouse HNSCC. C, ACTA2 expression results from bulk RNA sequencing data of HNSCC from the TCGA database. Data are presented as range and mean ± SEM, analyzed by the Wilcoxon test. D, Survival analysis of patients with HNSCC shows that high ACTA2 expression is not associated with overall survival rates. Fig. 4. The macrophage clearance leads to an increase in the CD8+ T/CD4+ T ratio. A, Expression changes of macrophage markers and apCAF-related markers in THP1 cells after stimulation with different concentrations of MCSF for 6 h. B and E, Flow cytometry dot plots and quantitative results showing the changes in the number of FAP+ cells after macrophage depletion. C and F, Flow cytometry dot plots and quantitative results showing the changes in the number of CD4+ T cells following macrophage depletion. D and G, Flow cytometry dot plots and quantitative results showing the changes in the number of CD8+ T cells after macrophage depletion. Statistics are shown in mean ± SD (F) accessed by the unpaired t test. *, P &lt; 0.05; **, P &lt; 0.01; ***, P &lt; 0.001; ns, nonsignificant, respectively. Fig. 5. Co-inoculation of apCAFs with SCC7 results in an increased number of apCAFs in the tumor tissue. A, Gross morphology and tumor tissue images of mice 14 days after inoculation with SCC7 alone or co-inoculation of SCC7 and apCAFs. B, Flow cytometry dot plots showing the changes in the number of FAP+ cells in tumor tissue 14 days after inoculation with SCC7 alone or co-inoculation with SCC7 and apCAFs. C, Quantitative bar chart of tumor volume in mice from both groups. D, Quantitative bar chart showing the changes in the number of FAP+ cells in tumor tissue from both groups. Statistics are shown in mean ± SD (F) accessed by the unpaired t test. *, P &lt; 0.05; **, P &lt; 0.01; ***, P &lt; 0.001; ns, nonsignificant, respectively.