Supplementary Figure 3: Increased activation of Bach2^−/– cells is antigen dose dependent and proliferation independent.

a, CD44 and CD62L expression 4 days after stimulation of naïve WT and KO OT-I CD8⁺ T cells with indicated concentrations of cognate peptide ligand (SL9) in the presence of congenically distinct feeder splenocytes. Numbers in gates indicate percentages. Data are representative of 2 independently repeated experiments. Naïve cells for assays were isolated by flow cytometric sorting. b, CFSE dilution and CD62L expression amongst CFSE-labelled WT and KO naïve CD8⁺ T cells stimulated in vitro for 3 days. c, Expression of CD69 and CD25 at day 4 following in vitro stimulation of naïve WT and KO CD8⁺ T cells with platebound anti-CD3 and anti-CD28 in the presence of IL-2. d, CFSE dilution and Annexin V staining on CFSE-labelled WT and KO naïve CD8⁺ T cells stimulated in vitro for 3 days with platebound anti-CD3 and anti-CD28 antibodies in the presence of 100 IU IL-2. Numbers in gates indicate percentages. e, Naïve WT and KO CD8⁺ T cells were stimulated with anti-CD3 and anti-CD28 antibodies in the presence of 100IU IL-2 and harvested for analysis at day 4. Expression of the indicated proteins was measured by SDS-PAGE and western blotting. Data are representative of 2 independently repeated experiments.