Fig. 3.

The release of THREDs depends on mtROS, and their inhibition is associated with reduced production of inflammatory cytokines. (A) Mouse naïve CD4⁺ T cells were left nonactivated or activated as depicted with or without mtROS inhibitor SKQ1. After 24 h, cells were analyzed by confocal microscopy with staining for DNA (SYTO13, green), CD4 (blue), and MitoTracker (red). (Scale bars: 20 μm.) (B and C) Parallel cultures were analyzed for gene expression (B) and cytokine release (C) after 72 h of stimulation. In B, expression of target genes is presented in arbitrary units (AU), as a percentage of the housekeeping gene Gapdh ± SD. In C, cytokine production is expressed as mean ± SEM of quintuplicate wells pooled and tested in triplicate. *P < 0.05; **P < 0.005; ***P ≤ 0.001. P values were determined by the two-tailed unpaired t test. The data in A–C are representative of three independent experiments.