Figure EV3. Differences in TCR repertoires between naïve and VM T cells, related to Fig 3 .

• A
  Alternative gating strategy for the same samples as shown in Fig 3B–D (T cells from Vβ5 mice). A representative experiment out of eight in total.
• B, C
  The expression of CD5 on CD8⁺ K^b‐OVA‐4mer⁺ CD8⁺ Vα2⁺ or Vα8.3⁺ cells or total CD8⁺ K^b‐OVA‐4mer⁺ isolated from LN and the spleen was determined by flow cytometry. Relative MFI CD5 levels were quantified (CD5 levels on CD8⁺ K^b‐OVA‐4mer⁺ Vα8.3⁺ were arbitrarily set as 1). Mean + SD, n = 3 independent experiments. Statistical significance was determined using two‐tailed one‐value t‐test.
• D
  TCRα sequences from Fig 3F were analyzed for TRAJ usage. Means + SEM. n = 4 independent experiments. Statistical significance was determined by chi‐square test.
• E
  Schematic representation of the generation and analysis of retrogenic monoclonal T‐cell subsets.
• F
  Frequency of VM T cells among monoclonal populations expressing TCR clones V14‐C2, V14‐C7, and OT‐I generated as in Fig 3G.
• G
  CD5 levels on naïve monoclonal T cells expressing V14‐C2, V14‐C7, and OT‐I. Representative mice out of 9–14 in total from two to four independent experiments.