Journal: Journal of Translational Medicine

Article Title: CD226 knockout alleviates high-fat diet induced obesity by suppressing proinflammatory macrophage phenotype

doi: 10.1186/s12967-021-03150-4

Figure Lengend Snippet: HFD increased CD226 expression in ATMs. A Comparison of soluble CD226 levels in sera of normal, overweight and obese individuals. Plots show individual data values obtained (79 individuals in total). Horizontal black lines represent median ± IQR. Differences between groups were determined by Kruskal–Wallis test. B The expression profiles of CD226 on myeloid cells, including monocytes, macrophages, and neutrophils. C , D FACS analysis of CD226 expression on macrophages in the spleen and epididymal SVF of C57BL mice fed with chow or a HFD for 16 weeks ( n = 3). E Immunofluorescence staining for F4/80 (red) and CD226 (green) in the epididymal fat of mice fed with chow or a HFD for 16 weeks ( n = 3). Scale bar means 100 μm. Data represent mean ± SEM. Differences between groups were determined by two-tailed, unpaired Student’s t -test. * P < 0.05, ** P < 0.01

Article Snippet: The following antibodies were used for western blotting and immunofluorescence: anti-PPAR-γ mAb (clone: C26H12, No: 2435), anti-AKT mAb (clone: 40D4, No: 2920), anti-p-AKT Ser473 mAb (clone: D9E, No: 4060), anti-p-FOXO1 Ser256 mAb (clone: E1F7T, No: 84192), and anti-perilipin-1 mAb (clone: D1D8, No: 9349), purchased from Cell Signaling Technology (Danvers, MA, USA); anti-p-VAV1 Tyr174 mAb (clone: 28.Tyr 174, No: sc-135785), anti-VAV1 mAb (clone: D-7, No: sc-8039), anti-FOXO1 mAb (clone: C-9, No: sc-374427), and anti-β-ACTIN mAb (clone: 2Q1055, No: sc-58673), purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA); anti-F4/80 mAb (clone: CI:A3-1, No: ab6640), purchased from Abcam Inc. (Cambridge, Cambridgeshire, UK); and CD226 pAb (No: 50232-M08H), purchased from Sino Biological Inc. (Beijing, China).

Techniques: Expressing, Comparison, Immunofluorescence, Staining, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: CD226 knockout alleviates high-fat diet induced obesity by suppressing proinflammatory macrophage phenotype

doi: 10.1186/s12967-021-03150-4

Figure Lengend Snippet: Deficiency of CD226 regulated macrophage M1/M2 polarization. A mRNA levels of M1/M2-type cytokines, Ccl3 , Cxcl14 , Cxcl16 , and PPAR-γ in LPS-stimulated peritoneal macrophages from WT and CD226KO mice (n = 3). B Glycolytic activity of LPS-stimulated peritoneal macrophages from WT and CD226KO mice. Combined data from three independent experiments (n = 3). C ROS production in LPS-stimulated peritoneal macrophages from WT and CD226KO mice (n = 3). Data represent mean ± SEM. Differences between groups were determined by unpaired Student’s t -test. * P < 0.05, ** P < 0.01

Article Snippet: The following antibodies were used for western blotting and immunofluorescence: anti-PPAR-γ mAb (clone: C26H12, No: 2435), anti-AKT mAb (clone: 40D4, No: 2920), anti-p-AKT Ser473 mAb (clone: D9E, No: 4060), anti-p-FOXO1 Ser256 mAb (clone: E1F7T, No: 84192), and anti-perilipin-1 mAb (clone: D1D8, No: 9349), purchased from Cell Signaling Technology (Danvers, MA, USA); anti-p-VAV1 Tyr174 mAb (clone: 28.Tyr 174, No: sc-135785), anti-VAV1 mAb (clone: D-7, No: sc-8039), anti-FOXO1 mAb (clone: C-9, No: sc-374427), and anti-β-ACTIN mAb (clone: 2Q1055, No: sc-58673), purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA); anti-F4/80 mAb (clone: CI:A3-1, No: ab6640), purchased from Abcam Inc. (Cambridge, Cambridgeshire, UK); and CD226 pAb (No: 50232-M08H), purchased from Sino Biological Inc. (Beijing, China).

Techniques: Activity Assay

Journal: Journal of Translational Medicine

Article Title: CD226 knockout alleviates high-fat diet induced obesity by suppressing proinflammatory macrophage phenotype

doi: 10.1186/s12967-021-03150-4

Figure Lengend Snippet: CD226 modulates the ATM polarization under a model of HFD-induced obesity. CD226 affects macrophage accumulation in adipose tissues and mediates macrophage M1 polarization. Proinflammatory M1-type ATMs induce the development of inflammation within adipose tissue and contribute to systemic inflammatory state, including fatty liver, by secreting proinflammatory cytokines through PPAR-γ-dependent signaling, which leads to the occurrence and progress of obesity

Article Snippet: The following antibodies were used for western blotting and immunofluorescence: anti-PPAR-γ mAb (clone: C26H12, No: 2435), anti-AKT mAb (clone: 40D4, No: 2920), anti-p-AKT Ser473 mAb (clone: D9E, No: 4060), anti-p-FOXO1 Ser256 mAb (clone: E1F7T, No: 84192), and anti-perilipin-1 mAb (clone: D1D8, No: 9349), purchased from Cell Signaling Technology (Danvers, MA, USA); anti-p-VAV1 Tyr174 mAb (clone: 28.Tyr 174, No: sc-135785), anti-VAV1 mAb (clone: D-7, No: sc-8039), anti-FOXO1 mAb (clone: C-9, No: sc-374427), and anti-β-ACTIN mAb (clone: 2Q1055, No: sc-58673), purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA); anti-F4/80 mAb (clone: CI:A3-1, No: ab6640), purchased from Abcam Inc. (Cambridge, Cambridgeshire, UK); and CD226 pAb (No: 50232-M08H), purchased from Sino Biological Inc. (Beijing, China).

Techniques:

Journal: iScience

Article Title: Early TRAIL-engagement elicits potent multimodal targeting of melanoma by CD34 + progenitor cell-derived NK cells

doi: 10.1016/j.isci.2023.107078

Figure Lengend Snippet:

Article Snippet: anti-DNAM-1 PE (REA1040) , Miltenyi Biotec B.V. , Cat# 130-117-489; RRID: AB_2727959.

Techniques: Virus, Recombinant, Multiplexing, Activity Assay, Software

Journal: PLoS ONE

Article Title: GMP-Compliant, Large-Scale Expanded Allogeneic Natural Killer Cells Have Potent Cytolytic Activity against Cancer Cells In Vitro and In Vivo

doi: 10.1371/journal.pone.0053611

Figure Lengend Snippet: (A) Cytotoxicity of expanded NK cells against various tumor cell lines was analyzed by 51 Cr-release assay with the indicated effector∶target (E∶T) ratio in triplicate. Cytotoxicity against normal PBMCs was also analyzed. The assay was performed two times with expanded NK cells from different donors, and representative data was presented. Each graph represents mean±SD. (B) Expression of HLA-class I, ULBP-1, ULBP-2, MIC-A/B, CD112 and CD155 was analyzed by flow cytometry in various tumor cell lines and normal PBMCs (solid lines). Grey histograms represent isotype controls. (C) Expanded NK cells were pre-incubated with blocking antibodies for DNAM-1, NKG2D, NKp30 and/or NKp44, and cytotoxicity was analyzed against SW480 or SNU398 cells by 51 Cr-release assay in triplicate. E∶T ratio was 10∶1. Percent inhibition of cytotoxicity was calculated as a percentage of the inhibition by the isotype control antibody. The assay was performed two times with expanded NK cells from different donors, and representative data are presented. Each bar graph represents mean+SD.

Article Snippet: For blocking experiments, NK cells were pre-incubated with 10 µg/mL anti-mouse IgG1κ (MOPC-21; BD Biosciences), 10 µg/mL anti-DNAM-1 (DX11; BD Biosciences), 2 µg/mL anti-NKG2D (149810; R&D systems), 2 µg/mL anti-NKp30 (P30-15; BioLegend) or 10 µg/mL anti-NKp44 (P44-8; BioLegend) for 30 min at 4°C, and 51 Cr-release assay was performed against SW480 and SNU398 at E∶T ratio of 10∶1.

Techniques: Release Assay, Expressing, Flow Cytometry, Incubation, Blocking Assay, Inhibition