Splenocytes and lymph node cells were collected from naïve, 2W1S-, B8R-, or gp33-peptide immunized WT and single TCR T cell mice, followed by magnetic tetramer enrichment of antigen-specific populations. Antigen-specific T cells were then stained and analyzed by flow cytometry. A, C, E) Representative plots of naïve (left) and immunized (right) WT and single TCR T cell mice showing 2W1S:Ab-, B8R:Kb-, or gp33:Db-specific populations respectively. B, D, F) Numbers antigen-specific T cells in naïve and antigen immunized WT and single TCR T cell mice. Each symbol represents one mouse. No statistically significant differences between the two genotypes were present at either of the time points (Student’s t-test). Flow plots are in Log10 fluorescence scale.