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: Extracellular staining was performed with BV421-coupled anti-TCRβ (H57-597, BD Biosciences), BUV395-labeled anti-CD4, APC-Cy7-conjugated anti-CD8 (eBioH35-17.2, Thermo Fisher Scientific), PE-Cy7-conjugated anti-CD44 (IM7, BD Biosciences), APC-coupled anti-CD62L (MEL-14, Thermo Fisher Scientific) and PE-conjugated anti-CD25 antibodies.
Techniques: Irradiation, Injection, Ex Vivo, Expressing, Two Tailed Test