Fig. 6

Neo-2/15 promotes ATP production and antitumor activity in CAR-NK cells via NRF1. a De novo motif analysis of XBP1s peaks at regions of open chromatin. “%target” is number of target sequences with motif over total target sequences; “%bkgd” (%background) is the number of background sequences with motif over total background sequences. p values calculated using binomial distribution in HOMER. b Genome browser images of XBP1s binding sites in regions of open chromatin in the vicinity of the NRF1 gene. c The expression of XBP1s and NRF1 of IL-2 or Neo-2/15 stimulated BBζ in tumor tissues detected by multicolor IF. CD56 (green), XBP1s (orange), NRF1 (white), or nucleus (DAPI; blue). d BBζ expanded with IL-2 or Neo-2/15 were collected from AsPC-1 grafts at specified time points post-injection. Expression of XBP1s and NRF1 were assessed using western blot. Killing potential (e) and ATP generation (f) of IL-2 or Neo-2/15 stimulated BBζ pretreated with 10058-F4 (60 μM), Toyocamycin (80 nM) or WRP139 (40 μM, NRF1 inhibitor) for 30 minutes and then co-cultured with AsPC-1 cells for 4 hours at E:T of 1:1. Data are presented as mean ± SD (*P < 0.05, **P < 0.01, ***P < 0.001, n = 4 in e and n = 5 in f). g Kaplan-Meier curve representing the percent survival of the experimental groups. Data are presented as mean ± SD (ns, not significant, **P < 0.01, ***P < 0.001, two-tailed log rank test, n = 6)