Cytokines/ Chemokines in Serum

1. Fig. S.1 A Cytokines/Chemokines in Serum B Cytokines/Chemokines in Tumor tissues Chemokine/Cytokine Quantification in Serum and in Tumor Microenvironment. Blood was taken from the animal and serum samples were kept in -80oC. The tumor was surgically removed and a tumor extract was prepared in T-per extraction buffer supplemented with protease inhibitor cocktail (Pierce, Rockford, IL). Ten milliliters of buffer was used per gram of tumor sample. Supernatant was kept frozen at -80oC. The concentrations of the indicated proteins were quantified in serum (A) and in tumor extract (B) with Milliplex MCYTOMAG-70K kit (Millipore) by the Luminex-200 assay.

2. Fig. S.2 Vector P8032+Isotype P8032+Anti-CXCR3 CD8+ CD8+ CD8+ Blocking of CXCR3 suppresses the infiltration of CD8 T cell . Mice with 5-day established CT26 tumors received anti-CXCR3 blocking or control mAb followed by P8032 vaccine. The same treatment was repeated twice at weekly intervals. On day 28, tumor sections were analyzed by immunofluorescence microscopy for the presence of CD8+ T cells (red). Fig. S.3 A Isotype Naive P8032 7DW8-5 P8032+7DW8-5 NK NKT Cell number CD69 B Isotype Naive P8032 7DW8-5 P8032+7DW8-5 DC DC Cell number CD86 CD80 Administration of 7DW8-5 results in activation of NKT cells, NK cells, and DC in mouse spleen. Mice were injected i.p. with 0.5 µg 7DW8-5 alone or with Salmonella vaccine. Mice were sacrificed after 48 h, spleens were harvested and single-cell suspensions were prepared by collagenase digestion to increase the yield of DCs. (A) NKT and NK cells were gated as TCRβ+/CD1d/PBS57 tetramer+ and TCRβ-/DX5+ cells, respectively and analyzed for CD69 expression. (B) DCs were gated as CD11c+ cells and analyzed for CD80 and CD86 expression. Shown are representative plots from 5 mice per group from one of two experiments.