other:
Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity.
Article Snippet: For isolation of tumor-infiltrating lymphocytes, tumor tissuewasminced into 1 to 2 mm pieces and digested with collagenase IV (1.25 mg/mL; #LS004188, Worthington), 0.1% soybean trypsin inhibitor (#T9128, Sigma), hyaluronidase (1 mg/mL; #LS002592, Worthington), and DNase I (100 mg/mL; #LS002007, Worthington) in complete DMEM for 30minutes at 37 C. Cell suspensionswere passed through a 70-mmcell strainer (Falcon) and resuspended in RPMImedia (Gibco).
MTT Assay:Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity.
Article Snippet: IL-6 R & D System Cat# 406-ML/CF Rec. .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022 .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022
In Situ:Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity.
Article Snippet: IL-6 R & D System Cat# 406-ML/CF Rec. .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022 .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022
Proximity Ligation Assay:Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity.
Article Snippet: IL-6 R & D System Cat# 406-ML/CF Rec. .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022 .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022
Enzyme-linked Immunosorbent Assay:Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity.
Article Snippet: IL-6 R & D System Cat# 406-ML/CF Rec. .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022 .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022
Extraction:Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity.
Article Snippet: IL-6 R & D System Cat# 406-ML/CF Rec. .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022 .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022
Cytotoxicity Assay:Article Title: Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity.
Article Snippet: IL-6 R & D System Cat# 406-ML/CF Rec. .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022 .. IL-1b R & D System Cat# 401-ML/CF STA-21 (STAT3 inhibitor) Santa Cruz Biotechnology Cat# SC-200757 Fludarabine (STAT1 inhibitor) Selleckchem Cat# S1491 79-6 (BCL6 inhibitor) Selleckchem Cat# S8250 Bortezomib EMD Millipore Cat# 504314 LPS Sigma Cat# L2630 Hoechst 33258 ThermoFisher Cat# H3569 Normal mouse serum Jackson ImmunoResearch Cat# 015-000-120 penicillin-streptomycin Gibco Cat# 10378016 phorbol 12-myristate 13-acetate (PMA) Sigma Cat# P8139 Ionomycin Sigma Cat# I3909 Brefeldin A Biolegend Cat# 420601 Critical commercial assays Thiazolyl Blue Tetrazolium Bromide (MTT) Millipore-Sigma Cat# M5655 DuolinkTM flowPLA Detection Kit Millipore-Sigma Cat# DUO94001 DuolinkTM In Situ PLA Probe Anti-Rabbit PLUS Millipore-Sigma Cat# DUO92002 DuolinkTM In Situ PLA Probe Anti-Mouse MINUS Millipore-Sigma Cat# DUO92004 DuolinkTM In Situ Wash Buffer Millipore-Sigma Cat# DUO82047 IgG Mouse ELISA Kit ThermoFisher Cat# 8850-40022 IgM Mouse ELISA Kit ThermoFisher Cat# 8850-47022 Tissue Extraction Reagent ThermoFisher Cat# FNN0071 LDH-Glo cytotoxicity assay kit Promega Cat# J2380 (Continued on next page) e3 Cell Reports Medicine 3, 100744, September 20, 2022
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