Journal: STAR Protocols

Article Title: High-throughput mass cytometry staining for deep phenotyping of human natural killer cells

doi: 10.1016/j.xpro.2022.101768

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Article Snippet: DNAM-1-171Yb (1:100) , Fluidigm , Cat# 3171013B.

Techniques: Recombinant, Blocking Assay, Staining, Lysis, Software, Antibody Labeling, Sterility, Microscopy, Spectrophotometry

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

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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: 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: