Extended Data Fig. 6: Intensified TCR stimulation induces high ROS production in TFH cells.
From: Selenium–GPX4 axis protects follicular helper T cells from ferroptosis
a, Purified naïve CD4+ T cells were stimulated with αCD3/CD28 in 2-Mercaptoethanol-deprived culture medium, and cytosolic ROS production was assessed by flow cytometry at 24 and 72 h after incubation (n = 3 for each dose). Data are represented as fold change by normalizing to no stimulation and representative of three independent experiments. b, c, Purified CD4+CD19–CD25–CD45RA–CXCR5high TFH cells from human tonsils were cultured with indicated cytokines or stimulators in the absence (b) or presence (c) of 3 µM RSL-3 in 2-Mercaptoethanol-deprived culture medium for 12 h. Lipid ROS levels and frequencies of 7AAD– viable cells were analysed by flow cytometry (n = 3). Data are representative of three independent experiments. d, Mice were subcutaneously immunised with NP-OVA in Alum. Flow cytometric analysis of cytosolic ROS production in CD44+CD25–CD19–CXCR5highPD-1high TFH and FAS+GL-7+B220+ BGC cells at day 10 post immunisation (n = 6). Each dot represents one mouse. Data are representative of two independent experiments with at least five mice per group. Data are presented as mean ± SEM (a–c) and analysed by two-sided Student’s t-test (a–c) or paired-sample t-test (d).
