Figure 6.

Myc protein expression correlates with effector functions

• A Lymph node CD8⁺ T cells were activated with N4 peptide (1 ng/ml) for 2 h, and GFP-Myc^neg and GFP-Myc^pos populations were FACS-sorted. Expression of IFNγ mRNA measured by qPCR relative to naïve unstimulated CD8⁺ T cells is shown. Data show mean + SEM of 3 biological replicates.
• B Flow cytometry data show GFP-Myc and IFNγ expression from spleen OT1 GFP-Myc^KI CD8⁺ T cells stimulated with 5 ng/ml Q4 (left panel) or T4 (middle panel) peptides for 24 h. Quadrant percentages are shown. The right panel shows the percentage of GFP-Myc^pos and GFP-Myc^neg CD8⁺ T cells expressing IFNγ protein. Data are from 3 biological replicates.
• C The fluorescence intensity of GFP-Myc and IFNγ expression from flow cytometry data modelled by linear regression in OT1 GFP-Myc^KI CD8⁺ T cells treated with 5 ng/ml Q4 for 24 h. Data are representative of 3 biological replicates.
• D Forward scatter and side scatter plots of CD69⁺ GFP-Myc^neg (upper panels) and CD69⁺ GFP-Myc^pos (lower panel) OT1 GFP-Myc^KI CD8⁺ T cells stimulated with 3.3 ng/ml Q4 or N4 peptides for 24 h. Data are representative of 2 biological replicates
• E, F GFP-Myc^KI CD8⁺ T cells were stimulated with CD3 antibody (60 ng/ml) for 24 h. Data are representative of 3 biological replicates. (E) Flow cytometry data show CD69 and GFP-Myc expression. (F) Forward and side scatter plots of CD69⁻ GFP-Myc^neg (left panel), CD69⁺ GFP-Myc^neg (middle panel) and CD69⁺ GFP-Myc^pos (right panel) populations.
• G CD4Cre⁻ Myc^fl/fl and CD4cre⁺ Myc^fl/fl lymph node cells were activated with CD3 (1 μg/ml) and CD28 (3 μg/ml) antibodies for 24 h. The data show forward and side scatter plots of CD4⁺ (upper panels) and CD8⁺ (lower panels) T cells, compared to unstimulated CD4Cre⁻ Myc^fl/fl cells. Data are representative of at least 3 biological replicates.