anti mouse cd8  (Proteintech)

Journal: PLOS Neglected Tropical Diseases

Article Title: NFATc1 regulates LAG3 + CD8 + T cells in the spleen of mice infected with Plasmodium yoelii NSM

doi: 10.1371/journal.pntd.0013605

Figure Lengend Snippet: (A) Representative spleen images of uninfected and infected mice. (B) Statistical analysis of mouse body weight and spleen weight. (C) Parasitemia was counted via tail vein blood smears under a microscope every 4 days post-Plasmodium infection (DPI). (D-F) Dynamic changes in the proportion and number of splenic CD8 + T cells detected by flow cytometry after infection. (G-K) Dynamic detection of Perforin, Granzyme B, CD107a, and IFN-γ secretion by splenic CD8 + T cells using flow cytometry. Each group included 4-6 mice, and experiments were repeated twice. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns : p > 0.05.

Article Snippet: Splenic lymphocytes were incubated with anti-mouse CD8a antibody-conjugated magnetic beads (Naive CD8a + T Cell Isolation Kit, Miltenyi Biotec) at 4°C for 30 minutes.

Techniques: Infection, Microscopy, Flow Cytometry

Journal: PLOS Neglected Tropical Diseases

Article Title: NFATc1 regulates LAG3 + CD8 + T cells in the spleen of mice infected with Plasmodium yoelii NSM

doi: 10.1371/journal.pntd.0013605

Figure Lengend Snippet: (A) RT-qPCR analysis of LAG3 mRNA levels in splenic immune cells from uninfected and infected mice. (B) Flow cytometric detection of LAG3 protein levels in splenic immune cells. (C) Violin plots showing LAG3 fluorescence intensity in splenic immune cells after Plasmodium infection. (D, E) Flow cytometry analysis of LAG3 distribution in splenic immune cells post-infection. (F) LAG3 mRNA levels in splenic CD8 + T cells (sorted by magnetic beads) from uninfected and infected mice, detected by RT-qPCR. (G) Dynamic changes in LAG3 protein expression in CD8 + T cells (detected by flow cytometry) in infected mice sacrificed every 4 days, with uninfected mice as controls. (H) spleen lymphocytes were isolated from uninfected mice and stimulated with lysates from 5-fold numbers of uRBC (5 × uRBC), 1-fold numbers of iRBC (1 × iRBC), 5-fold numbers of iRBC (5 × iRBC), or 10-fold numbers of iRBC (10 × iRBC), respectively, Anti-mouse CD28 monoclonal antibodies were plused, and LAG3 expression in CD8 + T cells was assessed by flow cytometry, 48 hours later. Each group included 4-6 mice, and experiments were repeated twice. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns : p > 0.05.

Article Snippet: Splenic lymphocytes were incubated with anti-mouse CD8a antibody-conjugated magnetic beads (Naive CD8a + T Cell Isolation Kit, Miltenyi Biotec) at 4°C for 30 minutes.

Techniques: Quantitative RT-PCR, Infection, Fluorescence, Flow Cytometry, Magnetic Beads, Expressing, Isolation, Bioprocessing

Journal: PLOS Neglected Tropical Diseases

Article Title: NFATc1 regulates LAG3 + CD8 + T cells in the spleen of mice infected with Plasmodium yoelii NSM

doi: 10.1371/journal.pntd.0013605

Figure Lengend Snippet: (A, B) Differential gene analysis of splenic LAG3 + CD8 + T cells vs. LAG3 - CD8 + T cells in infected mice using single-cell sequencing data, visualized by a volcano plot (A) and heatmap (B) . (C) Flow cytometric detection of co-expression of LAG3 and inhibitory molecules (PD1, TIM3, TIGIT, TGF-β) in CD8 + T cells. (D) Flow cytometry analysis of co-expression of LAG3 and activation markers (CD69, ICOS, CD25, CD38) in CD8 + T cells from uninfected and infected mice. (E) Flow cytometric detection of Perforin, Granzyme B, IFN-γ, and CD107a secretion in LAG3 + CD8 + T cells from uninfected and infected mice. (F) Effector and resting phenotypes of splenic LAG3 + CD8 + T cells vs. LAG3 - CD8 + T cells detected by flow cytometry. (G) Flow cytometric analysis of co-expression of LAG3 + CD8 + T cells and proliferation marker Ki67 before and after infection. Each group included 4-6 mice, and experiments were repeated three times. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns : p > 0.05.

Article Snippet: Splenic lymphocytes were incubated with anti-mouse CD8a antibody-conjugated magnetic beads (Naive CD8a + T Cell Isolation Kit, Miltenyi Biotec) at 4°C for 30 minutes.

Techniques: Infection, Sequencing, Expressing, Flow Cytometry, Activation Assay, Marker

Journal: PLOS Neglected Tropical Diseases

Article Title: NFATc1 regulates LAG3 + CD8 + T cells in the spleen of mice infected with Plasmodium yoelii NSM

doi: 10.1371/journal.pntd.0013605

Figure Lengend Snippet: (A) Violin plots showing NFATc1 expression in CD8 + T cells under uninfected vs. infected conditions, in splenic CD8 + T cells at 12 days post-infection, and in LAG3 + CD8 + T cells vs . LAG3-CD8 + T cells. (B) Flow cytometric detection of co-expression of LAG3 and NFATc1 on CD8 + T cells. (C) Predicted binding sites of transcription factor NFATc1 in the LAG3 promoter sequence via the JASPAR database. (D) Relative fluorescence intensity detected by dual-luciferase reporter assay in HEK 293T cells co-transfected with pGL3-LAG3 promoter plasmid and either pCDNA3.1(+)-NFATc1 plasmid or pCDNA3.1(+)-Vector plasmid for 48 hours. (E, F) qPCR detection of NFATc1 and LAG3 mRNA levels in HuT78 cells 48 hours after transfection with NFATc1-targeting siRNA. Each group included 4-6 mice, and experiments were repeated three times. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns : p > 0.05.

Article Snippet: Splenic lymphocytes were incubated with anti-mouse CD8a antibody-conjugated magnetic beads (Naive CD8a + T Cell Isolation Kit, Miltenyi Biotec) at 4°C for 30 minutes.

Techniques: Expressing, Infection, Binding Assay, Sequencing, Fluorescence, Luciferase, Reporter Assay, Transfection, Plasmid Preparation