Journal: Cell reports

Article Title: Integration of T helper and BCR signals governs enhanced plasma cell differentiation of memory B cells by regulation of CD45 phosphatase activity

doi: 10.1016/j.celrep.2021.109525

Figure Lengend Snippet: (A and B) CD45 phosphatase activity (A) and CD45 surface expression (B) in naive B cells (left panels) and MBCs (right panels), control (CTR) treated (filled histogram) or CD40L (open histograms). (C) CD45 phosphatase activity (upper panel, n = 13), panCD45 surface expression (middle panel, n= 7), and CD45 mRNA levels (lower panel, n = 3) of naive B cells (blue) and MBCs (red), CTR or CD40L treated. Graphs show pCAP-SP1 (upper panel) or CD45 (middle panel) MFI values relative to naive B cells. Lower graph: Quantitative real-time PCR of CD45 mRNA normalized to POLR2A expressed as RQ (relative quantity). (D and F) Activated BCR signaling kinases pSyk (D) and pErk (F) upon BCR cross-linking in naive B cells (left panel) or MBC (right panel) in CTR+vehicle (VEH) (filled histograms), CD40L+VEH (open histograms), or CD40L+CD45 inhibitor (open dotted histograms). (E and G) Graphs show pSyk (n= 9) (E) and pErk (n = 7) (G) MFI values relative to CTR naive B cells. (H and I) Real-time quantitative PCR analysis of SYK (H) and ERK (I) mRNA relative to POLR2A expressed as RQ (n = 3). (J) Purified human B cells were stimulated for 1 h at 37 ◦ C before CD45 activity was measured. Graph depicts % CD45 activity + B cells in CTR or CD40L-stimulated B cells (n= 4). Related to . **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: TaqManTM gene expression assay SYK Hs00895377_m1 , ThermoFisher , Cat# 4331182.

Techniques: Activity Assay, Expressing, Control, Real-time Polymerase Chain Reaction, Purification

Journal: Cell reports

Article Title: Integration of T helper and BCR signals governs enhanced plasma cell differentiation of memory B cells by regulation of CD45 phosphatase activity

doi: 10.1016/j.celrep.2021.109525

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: TaqManTM gene expression assay SYK Hs00895377_m1 , ThermoFisher , Cat# 4331182.

Techniques: Negative Control, Recombinant, Purification, Staining, Gene Expression, Lysis, Immunoprecipitation, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Blocking Assay, Cell Isolation, Software, Microscopy

Journal: Cell reports

Article Title: Antigen-Presenting Cell-Intrinsic PD-1 Neutralizes PD-L1 in cis to Attenuate PD-1 Signaling in T Cells

doi: 10.1016/j.celrep.2018.06.054

Figure Lengend Snippet: (A) Left: cartoon showing a cell culture assay in which PD-1 transduced Jurkat cells were stimulated with three types of antigen-loaded Raji B cells: (1) parental Raji cells that express neither PD-L1 nor PD-1, (2) Raji cells transduced with only PD-L1 (PD-L1+), and (3) Raji transduced with both PD-L1 and PD-1 (PD-L1+/PD-1+). On the right are FACS histograms showing PD-1 and PD-L1 surface expression of parental Jurkat cells, PD-1–mGFP transduced Jurkat (PD-1+) cells, parental Raji cells, PD-L1–mCherry transduced Raji (PD-L1+) cells, and Raji cells co-transduced with both PD-1–mGFP and PD-L1–mCherry (PD-L1+/PD-1+). a.u., arbitrary units. (B–D) Jurkat cells expressing PD-1–mGFP (PD-1+) were conjugated with Raji B cells (parental) (B), Raji cells transduced with only PD-L1–mCherry (PD-L1+) (C), or Raji cells co-transduced with PD-L1–mCherry and unlabeled PD-1 (PD-L1+/PD-1+) (D), as illustrated in the cartoon on the left (CD28 and B7 are omitted in this cartoon for simplicity). Shown on the right are confocal images of the cell conjugate acquired 2 min after cell-cell contact. mGFP and mCherry signals are shown as green and magenta, respectively. Scale bars, 10 µm. (E) Bar graph summarizing the interface enrichment indices (calculated as described in STAR Methods) of the three conditions shown in (B). Data are shown as mean ± SEM; n = 35 cells from three independent experiments. See also . (F) A representative western blot showing the levels ZAP70-Y493 phosphorylation and p85 (PI3K regulatory subunit) co-immunoprecipitated (IP) with CD28 from the lysates of the indicated Jurkat-Raji co-culture. Jurkat cells expressing PD-1–mGFP (PD-1+) were stimulated with Raji cells (parental), Raji transduced with PD-L1–mCherry (PD-L1+), or Raji cells co-transduced with PD-L1–mCherry and PD-1–mGFP (PD-L1+/PD-1+); the times at which the co-culture was lysed are indicated (STAR Methods). WCL, whole cell lysate. (G and H) Bar graphs summarizing immunoblots in (F), including pY493-ZAP70 immunoblot (G) and CD28 coIP p85 immunoblot (H). The optical density corresponding to each band was quantified by ImageJ and normalized to the conditions in which parental Raji cells were used as the APCs. Data are presented as mean ± SEM from three independent replicates. (I) Bar graph summarizing IL-2 levels in the medium of the indicated Jurkat-Raji co-culture, as described in (B)–(D), 24 hr after cell-cell contact. Data are presented as mean ± SEM from three independent measurements, with each run performed in triplicate.

Article Snippet: ZAP70 pY493 antibody , Cell Signaling Technology , Cat # 2704S; RRID: AB_2217457.

Techniques: Cell Culture, Transduction, Expressing, Western Blot, Phospho-proteomics, Immunoprecipitation, Co-Culture Assay

Journal: Cell reports

Article Title: Antigen-Presenting Cell-Intrinsic PD-1 Neutralizes PD-L1 in cis to Attenuate PD-1 Signaling in T Cells

doi: 10.1016/j.celrep.2018.06.054

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ZAP70 pY493 antibody , Cell Signaling Technology , Cat # 2704S; RRID: AB_2217457.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Cytotoxicity Assay, Software, Imaging

Journal: Cell reports

Article Title: T Cell-Intrinsic IRF5 Regulates T Cell Signaling, Migration, and Differentiation and Promotes Intestinal Inflammation

doi: 10.1016/j.celrep.2020.107820

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal phospho-Zap-70 (Tyr319)/Syk (Tyr352) , Cell Signaling Technology , Cat# 2701.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Sonication, DNA Library Preparation, Purification, Activation Assay, cDNA Synthesis, Software

Journal: EMBO Reports

Article Title: Functional BRI2-TREM2 interactions in microglia: implications for Alzheimer’s and related dementias

doi: 10.1038/s44319-024-00077-x

Figure Lengend Snippet: Reagents and tools.

Article Snippet: Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) , Cell Signaling Technology , 2717, RRID:AB_2218658.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Sequencing, Gene Expression, Blocking Assay, Western Blot, Isolation, Biomarker Discovery, Software, Imaging, Real-time Polymerase Chain Reaction

Journal: Cell Reports Medicine

Article Title: Oncolytic virus M1 functions as a bifunctional checkpoint inhibitor to enhance the antitumor activity of DC vaccine

doi: 10.1016/j.xcrm.2023.101229

Figure Lengend Snippet:

Article Snippet: Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb , CST , Catalog#: 2710.

Techniques: Control, Recombinant, Staining, Selection, Isolation, Red Blood Cell Lysis, Enzyme-linked Immunosorbent Assay, Reverse Transcription, SYBR Green Assay, RNA Sequencing, Software

Journal: Frontiers in Immunology

Article Title: Missing-in-Metastasis/Metastasis Suppressor 1 Regulates B Cell Receptor Signaling, B Cell Metabolic Potential, and T Cell-Independent Immune Responses

doi: 10.3389/fimmu.2020.00599

Figure Lengend Snippet: Key reagents table.

Article Snippet: Syk (D3Z1E) XP Rabbit mAb , 13198S , Cell Signaling Technology , Immunoblotting.

Techniques: Purification, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Recombinant, Western Blot, Microscopy

Journal: Immunity

Article Title: Immune Sensing of Cell Death through Recognition of Histone Sequences by C-Type Lectin-Receptor-2d Causes Inflammation and Tissue Injury

doi: 10.1016/j.immuni.2019.11.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: rabbit anti-Syk , Cell Signaling Technology , Cat#2712.

Techniques: Recombinant, Purification, Fractionation, Cell Culture, Luciferase, Lysis, Mass Spectrometry, Enzyme-linked Immunosorbent Assay, Software, Membrane

Journal: Cell reports

Article Title: Engineering chimeric antigen receptor neutrophils from human pluripotent stem cells for targeted cancer immunotherapy

doi: 10.1016/j.celrep.2022.111128

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Syk Rb mAb , Cell Signaling , Cat#13198T; RRID: AB_2687924.

Techniques: Virus, Recombinant, DNA Extraction, Staining, Enzyme-linked Immunosorbent Assay, Software, Imaging

Journal: iScience

Article Title: Aedes aegypti saliva modulates inflammasome activation and facilitates flavivirus infection in vitro

doi: 10.1016/j.isci.2023.108620

Figure Lengend Snippet:

Article Snippet: Syk (4D10) Mouse mAb , Cell Signaling , Cat# 80460 RRID: AB_2799953.

Techniques: Virus, Titration, Recombinant, Saline, Staining, Lysis, Extraction, Western Blot, Stripping, DC Protein Assay, ROS Assay, Enzyme-linked Immunosorbent Assay, Expressing, Software, Imaging, Spectrophotometry