Ova257 264 siinfekl

OVA_257–264 (SIINFEKL) was obtained from AnaSpec (Fremont, CA). Antibodies (Abs) against phospho-STAT3 (Tyr705) and STAT3 were from Cell Signaling Technology (Beverly, MA), and Ab against GAPDH was from Abcam (Cambridge, MA). Neutralizing Ab to G-CSF (MAB414) and isotype control (IgG1) were from R&D Systems (San Diego, CA). Neutralizing Ab to Gr-1 (RB6-8C5) and isotype control (IgG2b) were from BioXcell (West Lebanon, NH). Recombinant murine G-CSF and GM-CSF were from R&D Systems (San Diego, CA). Lentiviral expression of SOCS3 was generated as described (29 (link)).

OVA_257–264 (SIINFEKL) and gp100_25–33 (KVPRNQDWL) peptides were purchased from AnaSpec Inc (Fremont, CA). Recombinant human IL-2, IL-7 and IL-15 cytokines were kindly provided by the National Cancer Institute (NCI, MD). For fluorescence-activated cell sorting (FACS) analysis, mouse monoclonal antibodies to CD4 (GK1.5), CD8a (53–6.7), CD3 (HL3), IFN-γ (XMG1.2), CD25 (PC61), CD90.1 (OX-7), isotype control rat IgG2b (RTK4530), and IgG1 (RTK2071) were purchased from BioLegend (San Diego, CA); FoxP3 antibody (FJK-16s) was purchased from eBioscience (San Diego, CA). For Western blotting analysis, anti-human MDA-7/IL-24 antibody was purchased from GenHunter (Nashville, TN, USA); anti-mouse MDA-7/IL-24 antibodies were purchased from R&D Systems (Minneapolis, MN); phosphor-STAT1 (Y701), phosphor-STAT3 (Y705), total STAT1 and total STAT3 antibodies were purchased from Cell Signaling Technology (Davers, MA). Cucurbitacin I was purchased from Sigma Aldrich (St. Louis, MO). BrdU Flow Kit was purchased from BD Biosciences (San Jose, CA). Polybrene was purchased from Sigma Aldrich (St. Louis, MO). CytoTox 96^® Non-Radioactive Cytotoxicity Assay kits were purchased from Promega (Madison, WI). Cell Counting Kits 8 (CCK8) were purchased from Abcam (Cambridge, MA).