Fig. 5: Single-cell tumor immune microenvironment in PitNETs.

a UMAP plot showing the distribution of tumor immune microenvironment in PitNETs, colored by their cell types (n = 12,267 cells from 37 samples). b Bubble heatmap showing marker genes across 15 cell clusters through scRNA-seq (n = 12,267 cells from 37 samples). c Volcano plot showing differentially expressed genes between TAM_SPP1 versus TAM_IER2 (left), and TAM_SPP1 versus TAM_SEPP1 (right). A total of 1,367 TAMs were analyzed, including TAM_SEPP1 (n = 269 cells), TAM_SPP1 (n = 456 cells), and TAM_IER2 (n = 642 cells). Two-sided unpaired limma moderated t-test. d The pathway enrichment analysis of upregulated genes by TAM_SPP1. One-sided Fisher’s exact test. e Violin plot showing upregulated genes of TAM_SPP1 in TAM clusters. f Lollipop plot showing the spatial correlation coefficient between TAM_SPP1 cluster score and other dominant immune cell cluster scores (n = 35,347 spots from 20 samples). Spearman-correlation test. g Scatter plot showing the correlation between the infiltration of TAM_SPP1 and CAFs based on the average GSVA scores for each spatial transcriptomic (ST) sample (n = 20). Each circle represents one ST sample. The error band indicates a 95% confidence interval, the mean was used as the center for the error bars. Two-sided spearman-correlation test. h Spatial feature plots of signature score of TAM_SPP1 and CAFs in tissue sections. i Representative multiplex immunofluorescence staining of human PitNET tumors. DAPI (blue), CD68 (yellow), SPP1 (orange), αSMA (red), Ki67 (green), and tumor markers ACTH (ACTH1), TPIT (SCA5), and PIT-1 (TSH5) (magenta) are shown in individual and merged channels. Experiment was performed in three independent patients, each with three technical replicates. Scale bar, 100 µm. Source data are provided as a Source Data file for (c) and (e).