Fig. 2

The presence of SPP1⁺ Macs is positively associated with poor prognosis in HNSCC. A Volcano plot showing the upregulated genes and downregulated genes in myeloid cells. Adjusted p value < 0.05. B UMAP plots of all myeloid cells colored according to the expression level of SPP1. C Comparison of SPP1⁺ Mac percentages in paired adjacent normal tissues (n = 5) and tumor tissues (n = 5). D Representative IF staining of SPP1 (red) and various immune-associated cell types (green), including macrophages (CD68 +), CD4 + T cells, CD8 + T cells and B cells (CD20 +), and DAPI staining (blue) in human HNSCC tissues. Scale bar, 20 μm. E Representative IF staining of SPP1 (red) and CD68 (green) in paired adjacent tissues and HNSCC tissues. Scale bar, 20 μm. F PCR analysis of SPP1 expression in paired normal tissues (n = 74) and HNSCC tissues (n = 74). G Upregulated SPP1 expression compared with that in patients with stage I disease was correlated with advanced tumor stage. H ROC curves showing the ability of SPP1 to distinguish HNSCC tissues from adjacent normal tissues. I Kaplan‒Meier analysis of overall survival based on the SPP1 expression level in 74 pairs of HNSCC tissues. J Overall survival rate according to the SPP1⁺ Mac expression level in HNSCC tissues in a TCGA cohort. (**p < 0.01; ***p < 0.001; ****p < 0.0001)