Fig. 3.

Lymphocyte activation by PTPN3 inhibition was observed only in CD3⁺ T cells, not in CD3⁻ cells. a An FACS analysis of activated lymphocytes before selection. CD3⁺CD4⁺NKG2D⁻ cells (b) and CD3⁺CD8⁺NKG2D⁺ cells (c) were separately collected using microbeads. CD3⁻ (NK) cells (d) were collected using FACS sorting. e The migration ability of lymphocytes was analyzed by time-lapse imaging. The graph shows the moving distance of lymphocytes analyzed by the Image-Pro Analyzer software program. f The proliferation of FAR red-labeled lymphocytes was estimated by FACS. g The PTPN3 mRNA expression in CD3⁻ cells and CD3⁺ cells after activation was estimated by real-time RT-PCR. h The migration ability of non-activated lymphocytes was analyzed by the chamber method. i The cytotoxicity of non-activated lymphocytes was investigated. After co-culture of SCC cells and lymphocytes for 72 h, the absorbance of viable cancer cells was measured. j The PTPN3 mRNA expression in lymphocytes stimulated by allo-pancreatic cancer (SUIT-2) cell-pulsed mature DCs without using anti CD3 mAb was estimated by real-time RT-PCR. Similar results were obtained in two different healthy volunteers in all experiments. Data are presented as the mean ± SD. Lymphocytes and DCs from healthy volunteers were used in all experiments. *p < 0.05