51cr labeled yac 1 target cells  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: CD127 + Natural Killer Cells Represent a Distinct, Interleukin-15-Independent and Thymus-Independent Subset in Mice

doi: 10.3390/ijms27062667

Figure Lengend Snippet: Functional characterization of murine CD127 + NK cells. ( a ) Cytotoxic activity of sorted, freshly isolated CD127 + and CD127 − NK cells from the spleen and lymph nodes of adult C57BL/6 mice ( n = 5) was assessed against YAC-1 target cells using a 51 Cr-release assay at the indicated effector-to-target (E/T) ratios. CD127 + NK cells exhibited measurable cytotoxic activity; however, their lytic capacity was significantly lower than that of CD127 − NK cells at multiple E/T ratios. ( b ) Intracellular IFN-γ production by CD127 + and CD127 − NK cells following stimulation with anti-NK1.1 mAb (PK136). Representative flow cytometric plots are shown, with numbers in the quadrants indicating the percentage of cells in each region. Data are presented as mean ± SD. Statistical comparisons between CD127 + and CD127 − subsets were performed using an unpaired two-tailed Student’s t -test. * p < 0.05, ** p < 0.01. IFN-γ—interferon-γ; mAb—monoclonal antibody.

Article Snippet: Briefly, NK cell-mediated cytotoxicity was assessed by measuring 51 Cr-release following the incubation of freshly isolated, unstimulated effector cells with 51 Cr-labeled YAC-1 target cells (ATCC).

Techniques: Functional Assay, Activity Assay, Isolation, Release Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Aiolos restricts the generation of antigen-inexperienced, virtual memory CD8 + T cells in mice

doi: 10.1038/s41467-025-67540-8

Figure Lengend Snippet: a Heatmap showing differences in different cytokine receptors between WT and Aiolos-deficient T VM samples. b Schematic of the in vitro treatment of WT and Aiolos-deficient splenic CD8 + T cells, created in Microsoft PowerPoint. Bulk CD8 + T cells were isolated from the spleens of WT and Aiolos-deficient mice and treated with rmIL-12, rmIL-18, and IL-15C (IL-15/IL-15Rα complex) for 30 min (pY-STAT5 and pY-STAT4) or 16 h (IFN-γ, granzyme B and NKG2D) with protein transport inhibitor (PTI) added 4 h prior to harvesting the cells for flow cytometry analysis. Analysis of frequency (%) and median fluorescence intensity (MFI) fold change for ( c ) pY-STAT5 and ( d ) pY-STAT4 between WT and Aiolos-deficient T VM cells, relative to WT. Data representative of 5 independent experiments, n = 5/group, mean ± SEM; two-sided, unpaired Student’s t-test, *p ≤ 0.05 and **p ≤ 0.01. Frequency (%) and MFI fold change for e IFN-γ, f granzyme B, and g NKG2D between WT and Aiolos-deficient T VM cells, relative to WT. Data represents 6 independent experiments, n = 6/group for e , f ; and 5 independent experiments with n = 5/group for NKG2D. Data presented as mean ± SEM; two-sided, unpaired Student’s t-test, *p ≤ 0.05, ***p ≤ 0.001 and ****p ≤ 0.0001. h Percent (%) dead YAC-1 (target) cells when co-cultured with CD8 + T cells in the presence of cytokines (IL-12, Il-18, IL-15C), with or without anti-NKG2D antibody, data from 5 independent experiments with n = 5/condition. Data presented as mean ± SEM; two-way ANOVA with Tukey’s multiple comparisons test *p ≤ 0.05 and ****p ≤ 0.0001. Source data are provided as a file.

Article Snippet: The EL4 murine T cell lymphoma line (TIB-39) for transfection studies and YAC-1 cells (TIB-160) for cytotoxicity were acquired from the American Type Culture Collection (ATCC) and maintained in complete RPMI (RPMI media [catalog # 61870-036, Thermo Fisher Scientific] containing 10% FBS [catalog # 26140-079, Life Technologies] and 1% penicillin/streptomycin [catalog # 15140-122, Life Technologies]).

Techniques: In Vitro, Isolation, Flow Cytometry, Fluorescence, Cell Culture