Supplementary Figure S7 from Repolarization of Immunosuppressive Macrophages by Targeting SLAMF7-Regulated CCL2 Signaling Sensitizes Hepatocellular Carcinoma to Immunotherapy

(A-B) qPCR and ELISA analyses of CCL2 transcriptional and secretory levels in the indicated HCC cells. (C-D) Flow cytometry analysis of CD80 and CD206 expressions in THP-1-differentiated macrophages cocultured with the indicated HCC cells in the presence of recombinant CCL2 protein or CCL2 neutralizing antibody. (E-F) Chemotaxis assays showing the effect of CCL2 signaling on SLAMF7-regulated migration of macrophages using the indicated treatments. (G) Left: Gross appearance of the liver bearing orthotopic HCC tumors in the indicated treatment groups. Right: The tumor weight and tumor volume of each group (5 mice per group). (H) Flow cytometry assays of tumor infiltration proportions of CD11b+ F4/80+ MHC-II+ macrophages and CD11b+ F4/80+ CD206+ macrophages in the indicated treatment groups. *P<0.05, **P<0.01, and ***P<0.001, One-way ANOVA with a post hoc LSD test (or Student’s t test in part A-B). Experiments were repeated three time and results were present as the mean ± SD.