Journal: The Journal of Immunology Author Choice

Article Title: The Regulation of Nucleic Acid Vaccine Responses by the Microbiome

doi: 10.4049/jimmunol.2300196

Figure Lengend Snippet: Details of Ab reagents used in these experiments

Article Snippet: Ab target Conjugation Clone Manufacturer Product Number Concentration OVA-specific IgG1 None L71 Chondrex 7093 OVA-specific IgG2b None 4B436 Chondrex 7096 OVA-specific IgG2c None 3E3A9 Chondrex 7109 Mouse IgG HRP Poly4053 BioLegend 405306 1:3000 dilution Mouse IgG1 HRP — Invitrogen A10551 0.5 μg/ml Mouse IgG2b HRP — Invitrogen M32507 0.5 μg/ml Mouse IgG2c HRP — Invitrogen PAI-29288 0.5 μg/ml Mouse CD16/32 None 93 SouthernBiotech, BioLegend 1630-01 1 μg/ml Mouse CD3 BUV395 145-2C11 BD Biosciences 563565 2 μg/ml Mouse CD4 PerCP-eFluor710 RM4-5 Invitrogen 46-0042-82 1 μg/ml Mouse CD8 BUV737 53-6.7 BD Biosciences 612739 1 μg/ml Mouse IFN-γ Alexa Fluor 488 XMG1.2 BioLegend 505813 2 μg/ml Mouse TNF-α Pacific Blue MP6-XT22 BioLegend 506318 2 μg/ml Mouse IL-17 PE TC11-18H10 BioLegend 506904 1 μg/ml Mouse IL-4 PE-CF594 11B11 BioLegend 504132 1 μg/ml Mouse IL-10 APC JES5-16E3 BioLegend 505010 1 μg/ml Mouse CD11b BUV395 M1/70 BD Biosciences 563553 1 μg/ml Mouse CD11c FITC N418 BioLegend 117306 5 μg/ml Mouse Ly6G BV510 1A8 BioLegend 127633 1 μg/ml Mouse Ly6C BV785 HK1.4 BioLegend 128041 1 μg/ml Mouse CD103 PECy7 2E7 BioLegend 121426 1 μg/ml Mouse CD8a BUV730 53-6.7 BD Biosciences 612759 1 μg/ml Mouse XCR1 APCCy7 ZET BioLegend 148223 1 μg/ml Mouse CD301b EF660 11A10-B7 Invitrogen 50-3011-80 1 μg/ml Mouse CD64 BV605 X54-5/7.1 BioLegend 139323 1 μg/ml Mouse CD80 BV650 16-10AE BioLegend 104732 1 μg/ml Mouse CD86 PE PO3.1 eBioscience 12-0861-82 1 μg/ml Mouse MHC-II (I-A/I-E) AF700 M5/114.15.2 Invitrogen 565321-82 1 μg/ml CD45 BUV805 30-F112 BD Biosciences 748370 1 μg/ml CD107a FITC 1D4B BD Biosciences 553793 5 μg/ml CD4 PerCP-Cy5.5 GK1.5 BioLegend 100434 2 μg/ml CD8a Pacific Blue 53-6.7 BioLegend 100725 5 μg/ml CD3e BV605 145-2C11 BioLegend 100351 2 μg/ml IL-4 BV786 11B11 BD Biosciences 564006 2 μg/ml IL-2 APC JES6-5H4 BD Biosciences 554429 2 μg/ml IFN-γ AF700 XMG1.2 BD Biosciences 557998 2 μg/ml IL-17 APC-Cy7 TC11-18H10.1 BioLegend 506940 2 μg/ml IL-5 PE TRFK5 BioLegend 504304 2 μg/ml TNF-α PE-Cy7 MP6-XT22 BD Biosciences 557644 2 μg/ml Open in a separate window Details of Ab reagents used in these experiments

Techniques: Conjugation Assay, Concentration Assay

Journal: The Journal of Immunology Author Choice

Article Title: The Regulation of Nucleic Acid Vaccine Responses by the Microbiome

doi: 10.4049/jimmunol.2300196

Figure Lengend Snippet: Flow cytometry profiling of innate immune response to mRNA-LNP in GF and SPF mice. (A) Representative flow cytometry and graphical summaries of Ly6Chi monocyte gating and monocyte CD11c versus MHC class II expression in PBS-treated or mRNA-LNP-immunized GF and SPF mice. (B) Gating strategy to identify neutrophils and Ly6C+, CD64+, and CD64−Ly6C− DC/macrophage populations among splenocytes from PBS-treated or mRNA-LNP-immunized GF and SPF mice. (C) Graphical summaries of neutrophil and Ly6C+, CD64+, and CD64− Ly6C− DC/macrophage proportions in PBS-treated or mRNA-LNP–immunized mice. (D) Median fluorescent intensity (MFI) for CD86 and CD80 expression on the specified populations of immune cells from the spleens of GF or SPF mice 24 h after immunization with mRNA-LNP vaccine or PBS control. Each data point on graphs represents a single mouse (n = 3 per PBS group, n = 4 per mRNA-LNP group from one experiment). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA.

Article Snippet: Ab target Conjugation Clone Manufacturer Product Number Concentration OVA-specific IgG1 None L71 Chondrex 7093 OVA-specific IgG2b None 4B436 Chondrex 7096 OVA-specific IgG2c None 3E3A9 Chondrex 7109 Mouse IgG HRP Poly4053 BioLegend 405306 1:3000 dilution Mouse IgG1 HRP — Invitrogen A10551 0.5 μg/ml Mouse IgG2b HRP — Invitrogen M32507 0.5 μg/ml Mouse IgG2c HRP — Invitrogen PAI-29288 0.5 μg/ml Mouse CD16/32 None 93 SouthernBiotech, BioLegend 1630-01 1 μg/ml Mouse CD3 BUV395 145-2C11 BD Biosciences 563565 2 μg/ml Mouse CD4 PerCP-eFluor710 RM4-5 Invitrogen 46-0042-82 1 μg/ml Mouse CD8 BUV737 53-6.7 BD Biosciences 612739 1 μg/ml Mouse IFN-γ Alexa Fluor 488 XMG1.2 BioLegend 505813 2 μg/ml Mouse TNF-α Pacific Blue MP6-XT22 BioLegend 506318 2 μg/ml Mouse IL-17 PE TC11-18H10 BioLegend 506904 1 μg/ml Mouse IL-4 PE-CF594 11B11 BioLegend 504132 1 μg/ml Mouse IL-10 APC JES5-16E3 BioLegend 505010 1 μg/ml Mouse CD11b BUV395 M1/70 BD Biosciences 563553 1 μg/ml Mouse CD11c FITC N418 BioLegend 117306 5 μg/ml Mouse Ly6G BV510 1A8 BioLegend 127633 1 μg/ml Mouse Ly6C BV785 HK1.4 BioLegend 128041 1 μg/ml Mouse CD103 PECy7 2E7 BioLegend 121426 1 μg/ml Mouse CD8a BUV730 53-6.7 BD Biosciences 612759 1 μg/ml Mouse XCR1 APCCy7 ZET BioLegend 148223 1 μg/ml Mouse CD301b EF660 11A10-B7 Invitrogen 50-3011-80 1 μg/ml Mouse CD64 BV605 X54-5/7.1 BioLegend 139323 1 μg/ml Mouse CD80 BV650 16-10AE BioLegend 104732 1 μg/ml Mouse CD86 PE PO3.1 eBioscience 12-0861-82 1 μg/ml Mouse MHC-II (I-A/I-E) AF700 M5/114.15.2 Invitrogen 565321-82 1 μg/ml CD45 BUV805 30-F112 BD Biosciences 748370 1 μg/ml CD107a FITC 1D4B BD Biosciences 553793 5 μg/ml CD4 PerCP-Cy5.5 GK1.5 BioLegend 100434 2 μg/ml CD8a Pacific Blue 53-6.7 BioLegend 100725 5 μg/ml CD3e BV605 145-2C11 BioLegend 100351 2 μg/ml IL-4 BV786 11B11 BD Biosciences 564006 2 μg/ml IL-2 APC JES6-5H4 BD Biosciences 554429 2 μg/ml IFN-γ AF700 XMG1.2 BD Biosciences 557998 2 μg/ml IL-17 APC-Cy7 TC11-18H10.1 BioLegend 506940 2 μg/ml IL-5 PE TRFK5 BioLegend 504304 2 μg/ml TNF-α PE-Cy7 MP6-XT22 BD Biosciences 557644 2 μg/ml Open in a separate window Details of Ab reagents used in these experiments

Techniques: Flow Cytometry, Expressing, Control

Journal: EMBO Molecular Medicine

Article Title: CXCR4 engagement triggers CD47 internalization and antitumor immunization in a mouse model of mesothelioma

doi: 10.15252/emmm.202012344

Figure Lengend Snippet: Representative IHC staining for F4/80, CD45R, CD3, and HMGB1 in two mouse mesotheliomas. Nuclei were counterstained with hematoxylin. Scale bar 20 µm. Representative immunofluorescence (IF) staining for F4/80 (red), CD86 (green, in the upper panel), and CD206 (green, in the lower panel) of mouse MM lesions. Scale bar 50 µm. Representative IHC staining for CD68, CD206, CD163, CD20, CD3, and HMGB1 in human sarcomatoid and epithelioid mesotheliomas. Scale bar 50 µm.

Article Snippet: For immunofluorescent staining, 6‐μm‐thick serial cryostat sections from mice tumor samples, immediately snap‐frozen in OCT after removal, were fixed with cold acetone for 10 min and co‐stained with the following Ab: rabbit anti‐mouse mAb to F4/80 (1:100, Biolegend), rat anti‐mouse mAb to CD206 (1:200 Bio‐Rad), and rat anti‐mouse mAb to CD86 (clone PO.3, 1:100, #04‐1527 Millipore).

Techniques: Immunohistochemistry, Immunofluorescence, Staining