Fig. 5

VSIG4⁺ TAMs regulating SPP1 signaling to recruit neutrophil and shaped the immunosuppressive microenvironment. (A) The differences of SPP1 signaling in cell-cell communication after VSIG4 knockout. (B) The proportions of tumor-infiltrating neutrophils (CD11b⁺ Ly6G⁺) were measured by flow cytometry in wild-type (n = 6) and VSIG4-KO (n = 6) mice bearing mATC-derived tumor. (C) Gene set enrichment analysis to evaluate the scores of indicated pathways in different TAM subpopulations in human ATC samples. (D) The correlation between VSIG4 and SPP1 in PDAC was retrieved from GEPIA database. (E) Pan-cancer analysis of the correlation between VSIG4 and SPP1 in myeloid cells using TISCH2 database. (F) The expressions of Spp1 after VSIG4 knockout in TAMs. (G-I) Co-culture of mATC with BMDM isolated from VSIG4-KO and WT mice to examine the expressions of Spp1 by RT-qPCR and western blotting (n = 3 for each group). Data are presented as mean ± S.D. (J-M) The mATC tumor-bearing mice were intraperitoneally treated with 10 mg/kg anti-VSIG4 and/or anti-SPP1 as single agent and combination therapy every 3 days, respectively. Tumors were isolated on day 35. The tumor growth curve, body weight, and tumor weight of each group were recorded. (N-S) The proportions of TILs in ATC tumor-bearing mice measured by flow cytometry in IgG isotype (n = 6) and anti-SPP1 groups (n = 6). Data are presented as mean ± S.E.M. *P < 0.05, **P < 0.01, ***P < 0.001