Fig. 3: IL-2 promoted cDNT survival through OX40 molecule.

a Statistical analysis of OX40 expression on cDNT with or without IL-2 for 24, 48, and 72 h based on flow cytometric analysis. b Relative changes in OX40 expression induced by IL-2 in cDNT were plotted as fold changes of B6 cDNT without IL-2 stimulation. c B6 cDNT and OX40 KO cDNT were stimulated with or without IL-2 for 24, 48, and 72 h, and apoptosis of the cells was detected through Annexin V staining. Representative flow cytometric image of Annexin V⁺ cDNT from each group. d Statistical analysis of Annexin V⁺ cells in each group at a different time point. e Relative changes of Annexin V⁺ cells with or without IL-2 in B6 cDNT were compared with that in OX40 KO cDNT at 24, 48, and 72 h. f Caspase 3/7 activation was determined in B6 cDNT and OX40 KO cDNT after stimulation with or without IL-2. g The relative changes of Caspase 3/7 activation with or without IL-2 in B6 cDNT were compared with the ratio in OX40 KO cDNT at 24, 48, and 72 h. h The percentages of EdU⁺ cells relative to the total numbers of B6 cDNT or OX40 KO cDNT with or without IL-2 was determined by flow cytometry in the indicated groups. i Statistical analysis of EdU⁺ cells in each group at different time points. j Relative changes of EdU⁺ cells with or without IL-2 in B6 cDNT were compared with that in OX40 KO cDNT at 24, 48, and 72 h. k The B6 cDNT and OX40 KO cDNT were stimulated with IL-2 or without IL-2, and Alamar Blue was incubated during the stimulation. The proliferations of the cells were detected at the absorbance of 570 nm at different time points. ^#P < 0.05, IL-2 + B6 cDNT versus B6 cDNT; ^*P < 0.05, IL-2 + B6 cDNT versus IL-2 + OX40 KO cDNT. l A total of 5 × 10⁶ B6 cDNT or OX40 KO cDNT were adoptively transferred into B6D2F1 recipient mice by tail vein injection in the presence or absence of IL-2/Fc protein. After 3 days, cDNT were analyzed by flow cytometry for proliferation and apoptosis with BrdU and Annexin V detection. Data are representative of three experiments with similar results. The data are presented as the mean ± SD, n = 5 in each group. *p < 0.05, **p < 0.01, NS no significance