Fig 2: (A) H&E staining results of HER2+ BRCA tissues confirmed the negative relation between TILs infiltration and S100A9/LDHA intensity (10 cases for each subgroup. Typical representative was chosen for presentation. Scale bar = 100 μm; **, p < 0.01. TILs were pointed out by green arrows). (B) IHC staining results of HER2+ BRCA tissues revealed that CD3+/CD4+/CD8+ TILs were enriched in the S100A9 sparse cases, while FOXP3+ TILs mostly infiltrated in S100A9 sufficient cases (10 cases for each subgroup. Typical representative was chosen for presentation. Scale bar = 100 μm; *, p < 0.05; **, p < 0.01). S100A9: S100 calcium-binding protein A9. BRCA: Breast cancer. HER2: Human epidermal growth factor receptor 2. PGK1: Phosphoglycerate kinase 1. LDHA: Lactate dehydrogenase A. ENO1: Enolase α. TILs: Tumour infiltrating lymphocytes. H&E: Haematoxylin and eosin. IHC: Immunohistochemistry. CD3: Cluster of differentiation 3 receptor. CD4: Cluster of differentiation 4 receptor. CD8: Cluster of differentiation 8 receptor. FOXP3, Forkhead/winged-helix transcription factor P3. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig 3: Double-label immunofluorescence staining results of HER2+ BRCA tissues confirmed that CD3+/CD4+/CD8+ TILs enriched in the S100A9 sparse cases, while FOXP3+ TILs mostly infiltrated in S100A9 sufficient cases (10 cases were involved). Typical representative was chosen for presentation (scalebar = 50 μm). S100A9: S100 calcium-binding protein A9. BRCA: Breast cancer. HER2: Human epidermal growth factor receptor 2. CD3: Cluster of differentiation 3 receptor. CD4: Cluster of differentiation 4 receptor. CD8: Cluster of differentiation 8 receptor. FOXP3: Forkhead/winged-helix transcription factor P3. TILs: Tumour infiltrating lymphocytes.

Fig 4: Analysis of immunohistochemistry staining for 30 HER2-positive breast cancer patients. (A,B) Representative pictures of immunohistochemistry staining of CCDC69, PPP1R16B, IL21R, FOXP3, CD86 and PD-L1 for HER2-positive breast cancer patients at Qingdao Central Hospital. (C–F) Scatter plot of correlation between four M1 MIRGs and Macrophages M1 maker CD86. (G–J) Scatter plot of correlation between four M1 MIRGs and PD-L1 (K–N) OS analysis of four M1 MIRGs in Her2-positive caner patients by KM Plotter (n = 30).

Fig 5: Violin plots of differently expressed of CCDC69, PPP1R16B, IL21R, and FOXP3 under differently expressed CTLA-4 and PD-1. (A–D) Violin plots of the CCDC69 expression under the different situations of CTLA-4 and PD-1 expression and p-value. (E–H) Violin plots of the PPP1R16B expression under the different situations of CTLA-4 and PD-1 expression and p-value. (I–L) Violin plots of the IL21R expression under the different situations of CTLA-4 and PD-1 expression and p-value. (M–P) Violin plots of the FOXP3 expression under the different situations of CTLA-4 and PD-1 expression and p-value.