Fig. 3

Representative images and quantification of multiplex immunohistochemical staining of tumours at baseline and after neoadjuvant treatment (post-NT). a–c Assessment of the expression levels of CD31 (endothelial cells, red) and NG2 (pericytes, green) in the lung cancer microenvironment before and after treatment, along with statistical analysis of the ratio of CD31⁺ to NG2⁺. a Representative images of the lung cancer microenvironment showing CD31 and NG2 staining. b Statistical analysis of the CD31⁺ to NG2⁺ ratio in baseline and post-neoadjuvant treatment samples (paired samples, n = 14). c Statistical analysis of the CD31⁺ to NG2⁺ ratio in baseline and post-neoadjuvant treatment samples within the pCR group (paired samples, n = 11) and the non-pCR group (paired samples, n = 3). d–f Perivascular CD4⁺ T cell count per μm² of vascular area. CD31 staining (red) represents the vascular area, while CD4 staining (pink) indicates the infiltration of CD4⁺ T cells. d Representative images of the lung cancer microenvironment showing CD31 and CD4 staining. e Statistical analysis of perivascular CD4⁺ T cell count per μm² of vascular area in baseline and post-neoadjuvant treatment samples (paired samples, n = 16). f Statistical analysis of perivascular CD4⁺ T cell count per μm² of vascular area in baseline and post-neoadjuvant treatment samples within the pCR group (paired samples, n = 13) and the non-pCR group (paired samples, n = 3). g–i Assessment of the expression levels of CD80 (pink) and CD11c (green) in the lung cancer microenvironment before and after treatment, along with statistical analysis of the infiltration of CD11c⁺CD80⁺ M1 macrophage. g Representative images of the lung cancer microenvironment showing CD80 and CD11c staining. h Statistical analysis of the CD11c⁺CD80⁺ M1 macrophage in baseline and post-neoadjuvant treatment samples (paired samples, n = 12). i Statistical analysis of the CD11c⁺CD80⁺ M1 macrophage in baseline and post-neoadjuvant treatment samples within the pCR group (paired samples, n = 9) and the non-pCR group (paired samples, n = 3). j Representative images of the lung cancer microenvironment showing PD-1 (pink) and CD8 (green) staining. k Statistical analysis of the PD-1⁺CD8⁺ T cell in baseline and post-neoadjuvant treatment samples within the pCR group (paired samples, n = 5) and the non-pCR group (paired samples, n = 3). l Representative images of the lung cancer microenvironment showing CD39 (pink) and CD8 (green) staining. m Statistical analysis of the CD39⁺CD8⁺ T cell in baseline and post-neoadjuvant treatment samples within the pCR group (paired samples, n = 6) and the non-pCR group (paired samples, n = 3). Data are mean ± SD. a–h Wilcoxon paired t test. No Significance (ns), ns: P > 0.05, *P < 0.05, and **P < 0.01