Extended Data Fig. 4 |. Adoptive transfer experiments with H1-depleted splenic T cells from H1cTKO;Vav-iCre mice.

a, Schematic diagram depicting adoptive T cell transfer to Rag1^KO mice, which are deficient in mature T and B cells, and to congenic CD45.1 mice. In brief, T cells were isolated from WT and H1cTKO;Vav-iCre mice by affinity Dynabead selection. In the case of transfer to Rag1^KO mice the cells were stained with CellTrace CFSE. 1 × 10⁶ cells were transplanted into recipient Rag1^KO or CD45.1 mice via injection in the retro-orbital sinus. After 1 week, mice were euthanized, and adoptive T cell transfer was assessed by flow cytometry of harvested spleens. T cell, spleen and syringe images are from Servier Medical Art. b, c, Cell division of CD8⁺ T cells transplanted in Rag1^KO mice was measured by flow cytometry (two representative traces of each genotype are displayed) after staining with CellTrace CFSE (g), and the percentages of CD4⁺ and CD8⁺ T cells that underwent <7 cell divisions were quantified (h, n = 6 mice per genotype). d, Number of CD4⁺ and CD8⁺ T cells in spleens of Rag1^KO recipient mice 7 days after transfer of donor WT or H1cTKO;Vav-iCre T cells. N = 6 mice per genotype. e, f, T cells were isolated from WT and cTKO spleens and lymph nodes by column separation. 1 × 10⁶ were injected into congenic C57BL/6 CD45.1 recipient mice. After 1 week, mice were sacrificed and adoptive T cell transfer was assessed by flow cytometry of donor CD45.2⁺ CD4⁺ and CD8⁺ T cells in harvested spleens. e, Representative flow cytometry plots and (f) quantification of donor-derived CD45.2⁺ CD4⁺ and CD8⁺ T cells in recipients of T cells from WT (n = 5) and H1cTKO;Vav-iCre (n = 5) mice is shown. Each data point represents the average of two recipients per donor. g, h, Cells were isolated from wild-type and H1cTKO;Vav-iCre spleens and LNs, and were stained for CD4, CD8, and with a fluorophore-conjugated anti-Fas antibody. Fas expression was assessed by flow cytometry. g, Representative histograms are shown alongside (h) quantification of Fas expression in CD4⁺ and CD8⁺ cells derived from WT (n = 4) and H1cTKO;Vav-iCre (n = 4) mice. Data are mean ± s.d.; unpaired t-test. ns, not statistically significant; * P ≤ 0.05; ** P < 0.01; *** P < 0.001.