CD4^dimCD8^bright T cells are a genuine subset of CD8⁺ T cells. (A-D) Gating strategy for live cells using side scatter area (SSC-A) and Aqua Live/Dead staining (A), followed by gating for singlet cells using forward scatter area (FSC-A) and forward scatter height (FSC-H; B), then for lymphocytes using SSC-A and FSC-A(C), and for T cells using CD3 or CD3 and CD8(D). (E) Gating for CD4⁻CD8⁺ and CD4^dimCD8^bright T cells using CD4 and CD8 based on a previous CD3 gate. (F) Representative isotype staining showing gate placement between CD4⁻CD8⁺ and CD4^dimCD8^bright T cells. (G-J) Visualization of CD4^dimCD8^bright T cells by confocal microscopy. PBMCs were immunostained using anti–CD4-PE (red; G) and anti-CD8-FITC (green; H), then coexpression was visualized by overlay (I). (J) Magnification of overlay in panel I. Arrows point to the same cell coexpressing CD4 and CD8 across the panels.