Figure 7

HMGN2, released by T-Ag activated CD8⁺T cells, transmembrane transported into tumor cells. HMGN2 protein and the supernatant of T-Ag activated CD8⁺ T cells were pre-labeled with FITC. Tca8113 cells were seeded at a density of 3 × 10⁴ per well in 24-well plates. After overnight growth, the cells were cultured in medium with FITC pre-labeled samples. (A) HMGN2 transport into tumor cells analyzed with fluorescence microscope. The three figures are the same area. (a) Light micrographs of Tca8113 cells. (b) Fluorescent micrographs of Tca8113 cells of Hoechst 33258 nuclear staining. (c) Fluorescent micrographs of FITC labeled HMGN2 protein distribution in Tca8113 cells. (B) The Tca8113 cells were analyzed with fluorescent microscope. (a, b, c) FITC pre-labeled HMGN2 as the positive control. (d, e, f) FITC pre-labeled CD8⁺ T cells supernatant. (a, d) Cells under a light microscope. (b, e) Cells under a fluorescent microscope. (c, f) Cells under a fluorescent microscope after cultured in medium with HMGN2 depleted samples. (C) The Tca8113 cells were analyzed with Flow Cytometry. (a) Untreated Tca8113 control. (b, d) Tca8113 cultured in medium with FITC labeled samples. (c, e) Tca8113 cells cultured in medium with HMGN2 depleted samples. Figures are representative of three independent experiments. (f) Error bars represent FITC positive rate (%) of Tca8113 cells after cultured in medium with FITC labeled or HMGN2 depleted sample for 1 hour. Data are represented as means ± SD of three independent experiments. *Significantly decreased compared to HMGN2 undepleted (p < 0.05).