Fig. 6: apCAFs and pro-inflammatory macrophages exhibit positive feedback regulation in the tumor microenvironment.

A Distribution (left) and proportion (right) of macrophages subclusters in tumors from control, apCAFs and MHC II^- CAFs groups (n = 2 mice/per group). B Expression levels of pro-inflammatory phenotype and pro-inflammatory phenotype related genes among various subclusters of macrophages. C Representative images of multiplex immunofluorescence (green: CD74; red: PDGFRA; white: CD68; purple: CD86) and distance from apCAFs or MHC II^- CAFs to pro-inflammatory macrophages. Statistical significance was determined using Mann–Whitney two-sided test. Multiplexed immunofluorescence assays are performed twice on tumor samples following assay optimization. D Spatial feature plot of pro-inflammatory macrophages signature in sample 1. ST: Spatial transcriptomics. E Correlation of signature score of apCAFs and pro-inflammatory macrophages in spatial transcriptomics data of sample 1. Correlation was evaluated using the two-sided Spearman rank correlation coefficient. F Correlation of signature score of apCAFs and pro-inflammatory macrophages in ACRG cohort (n = 300). Correlation was evaluated using the two-sided Spearman rank correlation coefficient. G Heatmap showing ligands activity and regulatory potential of the prioritized ligands in apCAFs to macrophages. H Schematic representation of apCAFs or MHC II^- CAFs and macrophages co-cultured experiments. Created in BioRender. Song, J. (2024) https://BioRender.com/r25u487. I The expression of pro-inflammatory genes and anti-inflammatory genes in macrophages after co-culture with apCAFs or MHC II^- CAFs (n = 3 in each group). J The proportion of pro-inflammatory macrophages (CD86⁺ cells and MHC II⁺ cells) in macrophages after co-culture with apCAFs or MHC II^- CAFs from five different patients. Error bars represent the mean ± SEM. Statistical significance was determined using one-way ANOVA, followed by a two-sided Dunnett’s test to compare the specific differences between the groups. Source data are provided as a Source Data file. K The prediction of ligand-receptor interaction activity from macrophage subclusters to apCAFs through “CellChat” algorithm. Pro-inflam Macro: pro-inflammation macrophages; Anti-inflam Macro: Anti-inflammation macrophages. L The prediction of ligand-receptor interactions activity between pro-inflammatory macrophages and apCAFs through “CellCall” algorithm. Pro-inflam Macro: pro-inflammation macrophages; Anti-inflam Macro: Anti-inflammation macrophages. M Schematic representation of control macrophages or pro-inflammatory macrophages and pan-CAFs co-cultured experiments. Pro-inflam Macro: pro-inflammation macrophages. Created in BioRender. Song, J. (2024) https://BioRender.com/h05n316. N The proportion of apCAFs in pan-CAFs from five different patients after co-culture with control macrophages or pro-inflammatory macrophages. Error bars represent the mean ± SEM. Statistical significance was determined using unpaired two-sided t-test. Source data are provided as a Source Data file. Pro-inflam Macro: pro-inflammation macrophages.