Figure 3

PHA-induced HMGN2 was mainly expressed in CD8⁺T cell population. PBMCs were seeded at a density of 1 × 10⁷ per well in 6 well plates and stimulated with 20 μg/ml PHA for 72 hours. Cells were collected and stained with CD3-FITC and CD8-PE. CD3⁺CD8⁺T cells (Gate R1), CD3⁺CD8^-T cells (Gate R2) and CD3^- mixed cells (Gate R3) were purified with MoFlo XDP sorter. 4 × 10⁶ CD3⁺CD8⁺T cells, CD3⁺CD8^-T cells and CD3^- mixed cells were incubated in 24-well plates respectively for 5 days. (A) The supernatants were collected and HMGN2 concentrations were analyzed by ELISA. (B) The cells were removed and intracellularly stained for HMGN2. Figures are representative of three independent experiments. (C) Error bars represent HMGN2 intracellular expression positive rate (%) in CD8⁺ T cells populations. Data are represented as means ± SD of three independent experiments. *Significantly higher compared to CD8^-CD3⁺ and MIX cells (p < 0.05).