Supplementary Tables S1-S8. Supplementary Figures S1-S12. from The Prognostic Role of Macrophage Polarization in the Colorectal Cancer Microenvironment

Abstract

<p>Supplementary Table S1. Analyses of MSI, DNA methylation, KRAS, BRAF, and PIK3CA mutations, and neoantigen load. Supplementary Table S2. List of antibodies and fluorophores used in the immunofluorescence procedure. Supplementary Table S3. Statistical methods. Supplementary Table S4. Macrophage density, M1-like macrophage density, M2-like macrophage density, and M1:M2 macrophage density ratio in overall tissue regions and patient survival with inverse probability weighting (IPW). Supplementary Table S5. Densities of macrophage populations defined by positivity for single polarization marker in tumor intraepithelial and stromal regions and patient survival with inverse probability weighting (IPW). Supplementary Table S6. Clinical, pathological, and molecular characteristics of colorectal cancer cases according to year of diagnosis. Supplementary Table S7. Macrophage densities and M1:M2 macrophage density ratio in tumor intraepithelial and stromal regions and patient survival in strata of MSI status with inverse probability weighting (IPW). Supplementary Table S8. Macrophage densities and M1:M2 macrophage density ratio in tumor intraepithelial and stromal regions and patient survival in strata of year of diagnosis with inverse probability weighting (IPW). Figure S1. Comparison of staining patterns between multiplex immunofluorescence and standard immunohistochemistry. Figure S2. Flowchart of the cyclic immunofluorescence procedure. Figure S3. Analysis of immunofluorescence slides to quantify tumor intraepithelial and stromal macrophage densities. Figure S4. Macrophage densities across 10 TMAs. Figure S5. Fluorescence signal intensities across 10 TMAs. Figure S6. t-SNE analysis of a random sample of 0.5% of all cells based on fluorophore signal intensities shows no clear clustering according to the TMAs. Figure S7. Core-to-core correlation of macrophage densities in two randomly chosen cores of tumors with two or more cores. Figure S8. Inverse probability weighting-adjusted Kaplan-Meier survival curves of colorectal cancer survival according to the ordinal quartile categories (Q1-Q4) of intraepithelial (A) and stromal (B) macrophage densities. Figure S9. Forest plots of inverse probability weighting-adjusted Cox regression models of colorectal cancer specific survival according to the densities of intraepithelial and stromal M1-like polarized macrophages, with M1-like macrophages defined using different cut-offs (10-50%) of the M1-end of the macrophage polarization (M1:M2) index distribution. Figure S10. Forest plots of inverse probability weighting-adjusted Cox regression models of colorectal cancer specific survival according to the densities of intraepithelial and stromal M2-like polarized macrophages, with M2-like macrophages defined using different cut-offs (10-50%) of the M2-end of the macrophage polarization (M1:M2) index distribution. Figure S11. Forest plots of inverse probability weighting-adjusted Cox regression models of colorectal cancer specific survival according to the intraepithelial and stromal M1:M2-density ratio with M1-like and M2-like macrophages defined using different cut-offs (10-50%) of both tails of the macrophage polarization (M1:M2) index distribution. Figure S12. Densities of M1-like and M2-like macrophages in relation to histologic lymphocytic reaction patterns.</p>