Fig. 1.
IL-35 and IL-10 suppress lysophosphatidylcholine (LPC)-induced human aortic endothelial cell (HAEC) activation by inhibiting mitochondrial reactive oxygen species (mtROS), adhesion molecules (ICAM1 and VCAM1), and proinflammatory cytokine IL1B; IL-35 is more potent than IL-10 in suppressing LPC-induced ICAM-1 expression. HAEC were stimulated by LPC (10 μM) with or without IL-35/IL-10/TGF-β (10 ng/mL) for 1 h (A) or 18 h (B&C), mtROS measurement using flow cytometry (n = 4 in each group) (A), Human Endothelial Cell Biology RT2 Profiler PCR Array (B; 3 pooled samples in each group), and Western Blot (C) were performed. Of note, transforming growth factor-β (TGF-β) served as a ‘house-keeping anti-inflammatory cytokine” control (see our paper, PMID: 22438968). For all panels, data are expressed as mean ± SEM and data are representative of at least two independent experiments. ***p < 0.001, *p < 0.05. MitoSOX is a fluorogenic dye to specifically measure mtROS in live cells. LPC, lysophosphatidylcholine; HAEC, human aortic endothelial cells; mtROS: mitochondrial reactive oxygen species.