Fig. 6.

Spatial interaction of mtLC-MSC and EFGR-wt tumor cells in EGFR-mt NSCLC tissues. A–H Immunohistochemistry staining assessed EGFR-wt tumor cells. Total 23 patients with EGFR-mt NSCLC were included. Representative images showing EGFR-wt cells within mutation-specific EGFR tumor tissues (A); intratumoral composition of EGFR-wt and EGFR-mt tumor cells by case (B); paired comparison of intratumoral EGFR-wt vs. EGFR-mt (C) and EGFR-wt vs. EGFR-mt proportions among total cells (D); proportion of EGFR-wt cells in tumor cells (E) and in total cells (F) compared between TKI-sensitive and TKI-resistant groups (sensitive n=14; resistant n=9); representative images of S100A9, FASN and SOX2 expressed in TKI-sensitive and TKI-resistant tumors (G); and statistical these marker expression in EGFR-wt tumor cells (H). (I–M) Multiplex immunofluorescence staining of paired mtLC-MSCs and mtTF-MSCs (n = 23). Representative CD105+ mtLC-MSC in TKI-resistant and sensitive tumors (I); proportions of MSCs in paired tumor-free (TF) and lung cancer (LC) tissues (J); schematic diagram of spatial analysis, using each tumor as the core to calculate nearest distance to the MSCs (K); statistical summary of mean nearest distance, using EGFR-mt and wt tumor cells as the core (L); and stratification by sensitivity (M). N–Q Multiplex immunofluorescence staining of spatial association between mtLC-MSC subsets and EGFR-wt tumor cells (n = 5). Representative S100A9 and FASN-positive expression in mtLC-MSC (N); FASN expression in S100A+ mtLC-MSCs (O); distribution of mean nearest differences for S100A9 and FASN-positive and negative mtLC-MSC subsets shown as quantile (25th, median, 75th; ΔQ1/Δmedian/ΔQ3) (P); and as continuous percentile curves (Q). Statistical results were shown as mean ± SD except panel J, K and M (mean ± SEM). *P<0.05, **P<0.01, ***P<0.001, **** P < 0.0001. Wilcoxon signed-rank test (C, D, J, L and M), Mann–Whitney U test (E, F and H) or Student’s t-test (O and P)