pe-cy7 anti-foxp3  (Thermo Fisher)

Journal: Frontiers in Immunology

Article Title: Dual variants of uncertain significance in a case of hyper-IgM syndrome: implications for diagnosis and management

doi: 10.3389/fimmu.2025.1594636

Figure Lengend Snippet: Immunophenotyping of T cells and assessment of IKKb expression. (A) , Representative dot plots gated on CD3 + CD4 + cells, illustrating the percentage of Tregs co-expressing CD25 and FoxP3. (B) , Comparison of CD45RO+ frequency in CD4+ and in CD8+ T cells in patient and siblings. (C) , Representative histogram of flow cytometry showing normal cytokine production comparing the patient, siblings and two healthy donors (HDs).

Article Snippet: The cells were then washed, fixed/permeabilized using the eBioscience FoxP3 Transcription Factor Staining Buffer kit (ThermoFisher Scientific) and incubated overnight with V450 anti-CD4 and PE-Cy7 anti-FoxP3 (eBioscience) antibodies.

Techniques: Expressing, Comparison, Flow Cytometry

Journal: bioRxiv

Article Title: CMTM6 suppresses cell-surface expression of death receptor FAS in mice but not in humans

doi: 10.1101/2025.05.12.653457

Figure Lengend Snippet: A .Weight and cellularity of spleens isolated from 8-12-week-old WT and Cmtm6 -/- mice. B . Flow cytometry analysis of splenic T cells from 8-12-weeks-old WT and Cmtm6 -/- mice. T cells (gated as CD3 + ) were subdivided into conventional CD4 + (CD4 + , FOXP3 − ), Tregs (CD4 + , FOXP3 + ), and conventional CD8 + (CD8 + ). C . Conventional CD4 + T cells were further gated as naïve (CD4 + , FOXP3 − , CD44 − , CD62L + ) and memory (CD4 + , FOXP3 − , CD44 + , CD62L − ). D . Conventional CD8 + T cells were gated as naïve (CD8 + , CD44 − ), memory (CD8 + , CD44 + , CD49d + ), and antigen-inexperienced memory T (AIMT) (CD8 + , CD44 + , CD49d − ) cells. E-J . Surface FAS levels in the indicated subsets of splenic T cells. Data are presented as mean + SEM, n=8 mice per group. Two-tailed Mann-Whitney test. ns, not significant.

Article Snippet: Antibody against FoxP3 PE-Cy7 (#25-5773-80) was from Thermo Fisher Scientific.

Techniques: Isolation, Flow Cytometry, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: CMTM6 suppresses cell-surface expression of death receptor FAS in mice but not in humans

doi: 10.1101/2025.05.12.653457

Figure Lengend Snippet: A .Body weight and spleen-to-body ratio in 8-12-week-old WT and Cmtm6 -/- mice. B . Flow cytometry analysis of T cells isolated from peripheral lymph nodes of 8-12-weeks-old WT and Cmtm6 -/- mice. T cells (gated as CD3 + ) were subdivided into conventional CD4 + (CD4 + , FOXP3 − ), Tregs (CD4 + , FOXP3 + ), and conventional CD8 + (CD8 + ). C . Conventional CD4 + T cells were further gated as naïve (CD4 + , FOXP3 − , CD44 − , CD62L + ) and memory (CD4 + , FOXP3 − , CD44 + , CD62L − ). D . Conventional CD8 + T cells were gated as naïve (CD8 + , CD44 − ), memory (CD8 + , CD44 + , CD49d + ), and antigen-inexperienced memory T (AIMT) (CD8 + , CD44 + , CD49d − ) cells. E-J . FAS expression in the indicated subsets of T cells isolated from peripheral lymph nodes. Data are presented as mean + SEM, n=8 mice per group. Two-tailed Mann-Whitney test. ns, not significant.

Article Snippet: Antibody against FoxP3 PE-Cy7 (#25-5773-80) was from Thermo Fisher Scientific.

Techniques: Flow Cytometry, Isolation, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT or HP1α-deficient naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. a Representative dot-plots showing 21 days after engraftment the percentage of WT and HP1α KO TCR Vβ6 + CD4 + T cells producing IFN-γ and IL-17A. b – d Percentage of WT or HP1α KO TCR Vβ6 + CD4 + T cells producing IFN-γ (b), IL-17A ( c ) or both ( d ) 21 days after engraftment. e Volcano plot showing results of differential gene expression analyses between Treg that had been co-injected with WT or HP1α KO naive CD4 + T cells. Red and black dots represent genes with higher expression in Treg co-injected with HP1α KO or WT cells, respectively. Gray dots represent genes that failed to reach the FDR threshold of 0.05 and the absolute log2 fold change threshold of 1. f Absolute number of spleen cells 21 days after engraftment. Data are mean ± SD of three (Tconv) or seven (Tconv + Treg) biological replicates from two independent experiments. g Percentage of Tconv, defined as CD4 + CD45.1 - CD45.2 + H-2K d- Thy1.1 - , in the spleen 21 days after engraftment. Data are mean ± SEM of three independent experiments. h Percentage of allospecific Tconv, defined as CD4 + CD45.1 - CD45.2 + H-2K d- Thy1.1 - TCRVβ6 + , in the spleen 21 days after engraftment. Data are mean ± SEM of three independent experiments. i, j Percentage of CD73 + FR4 + ( i ) or Foxp3 + ( j ) cells among WT or HP1α KO TCR Vβ6 + CD4 + T cells, as determined in the spleen 21 days after engraftment. b – d and g – j Data show mean ± SEM of 3 independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. Source data are provided in the Source data file.

Article Snippet: For transcription factor expression analysis, cells were labeled with PE-Cy7-conjugated anti-Foxp3 antibodies using the Transcription Factor Staining Buffer Set (Thermo Fisher Scientific).

Techniques: Irradiation, Injection, Ex Vivo, Expressing

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: Lethally-irradiated B6 mice were grafted with a 1:1 mixture of B6 and B6D2F1 bone marrow and co-injected with WT (a-d and I, j) or HP1γ-deficient ( a–j ) naive CD4 + T cells alone or in the presence of ex vivo expanded WT Treg. a Representative dot-plots showing 21 days after engraftment the percentage of WT and HP1γ KO TCR Vβ6 + Tconv producing IFN-γ and IL-17A. b, c Percentage of WT or HP1γ KO TCR Vβ6 + Tconv producing IFN-γ ( b ) or IL-17A ( c ) 21 days after engraftment. d Percentage of CD73 + FR4 + cells among WT or HP1γ KO TCR Vβ6 + Tconv, as determined in the spleen 21 days after engraftment. e Percentage of cytokine-producing cells among HP1γ KO TCR Vβ6 + Tconv 21 days post-engraftment. The analysis was performed on total Tconv from mice injected with Tconv only, or on anergic (A) or non-anergic (N-A) Tconv from mice injected with Tconv and Treg. ( f ) Representative dot-plots showing 21 days after engraftment the percentage of HP1γ KO TCR Vβ6 + Tconv expressing PD-1 and TIGIT. g, h Percentage of PD-1 + , TIGIT + , LAG-3 + ( g ) or PD-1 + TIGIT + LAG-3 + ( h ) cells among HP1γ KO TCR Vβ6 + Tconv 21 days after engraftment. i Percentage of Foxp3 + cells among WT or HP1γ KO TCR Vβ6 + Tconv, as determined in the spleen 21 days after engraftment. j Percentage of spleen Foxp3 + cells among anergic or non-anergic WT and HP1γ KO TCR Vβ6 + Tconv 21 days after engraftment. b – d and I, j Data are represented as mean ± SEM of three independent experiments. Each symbol represents the mean of an experiment. P values were calculated using multiple unpaired t test with Holm-Šidák correction. e, g, h Floating bar charts represent the mean of the biological replicates as well as the minimum and maximum values. Each symbol represents individual biological replicates. P values were calculated using unpaired t test (two-tailed). Source data are provided in the Source data file.

Article Snippet: For transcription factor expression analysis, cells were labeled with PE-Cy7-conjugated anti-Foxp3 antibodies using the Transcription Factor Staining Buffer Set (Thermo Fisher Scientific).

Techniques: Irradiation, Injection, Ex Vivo, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Heterochromatic gene silencing controls CD4 + T cell susceptibility to regulatory T cell-mediated suppression in a murine allograft model

doi: 10.1038/s41467-025-55848-4

Figure Lengend Snippet: HLA-B7 - WT or HP1γ-deficient human naive CD4 + T cells were injected i.v . into sublethally-irradiated NSG mice with or without ex vivo expanded HLA-B7 + human Treg. Three weeks after injection, splenocytes were isolated and the xenogeneic T cell response was analyzed by flow cytometry. a Experimental model. b , c Following genome editing by CRISPR-Cas9, the expression level of HP1γ was determined in WT and HP1γ KO human CD4 + T cells. A representative western-blot ( b ) and normalized expression levels for each experiment ( c ) are shown. d Representative dot-plots showing the percentage of HLA-B7 - WT and HP1γ KO human Tconv producing IFN-γ and GM-CSF. e – g Percentage of HLA-B7 - WT and HP1γ KO human Tconv producing IFN-γ ( e ), GM-CSF ( f ) or Granzyme B ( g ). h Expression level of T-bet in HLA-B7 - WT and HP1γ KO human CD4 + T cells. i , j Percentage of HLA-B7 - WT and HP1γ KO human CD4 + T cells producing IL-10 (i) or expressing Foxp3 ( j ). Data show values for biological replicates from two ( e – j ) or three ( c ) independent experiments. Horizontal bars represent mean ± SD ( e – j ) or mean ± SEM ( c ). P values were calculated using two-tailed unpaired t tests. Source data are provided in the Source data file.

Article Snippet: For transcription factor expression analysis, cells were labeled with PE-Cy7-conjugated anti-Foxp3 antibodies using the Transcription Factor Staining Buffer Set (Thermo Fisher Scientific).

Techniques: Injection, Irradiation, Ex Vivo, Isolation, Flow Cytometry, CRISPR, Expressing, Western Blot, Two Tailed Test