Fig. 7

CCL8 secreted from APN KO BMSCs within EL-4 tumor recruits CD8⁺ T cells. A The population of infiltrated CD8⁺ T cells was analyzed by flow cytometry. B APN KO BMSCs were placed in the bottom chamber, splenocytes from EL-4-bearing WT mice were placed in the upper chamber of the transwell plate, and the population of migrated CD8⁺ T cells in the bottom chamber was analyzed by flow cytometry. C Cell lysates from the BMSCs were used to measure the levels of various chemokines. All dots were quantified using ImageJ software, and bar graphs were used for quantitative data. D The mRNA levels of the indicated chemokines were measured using RT-PCR and real-time PCR. E The level of CCL8 in cell culture supernatant with or without the treatment of vitexin (20 μM) in WT and APN KO BMSCs was measured. F WT BMSCs were placed in the bottom chamber of the transwell plate with or without CCL8 (10 ng/ml). Splenocytes from EL-4-bearing WT mice were placed in the upper chamber of the transwell plate and migrated CD8.⁺ T cells in the bottom chamber were analyzed by flow cytometry. All images were quantified using ImageJ software, and bar graphs were used for quantitative data. All data were expressed as the mean ± SD from at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001