Figure 10: Preeclamptic-like cytokine cocktails strongly inhibit ATP-stimulated Ca²⁺ bursts and are partially rescued by CLA in the high VEGF₁₆₅ condition.

Cocktails corresponding to Normal (0.5ng/ml VEGF₁₆₅, 0.03ng/ml TNFα, 0.1ng/ml IL-6) or PE ((L)PE: 0.1ng/ml VEGF₁₆₅, 0.5ng/ml TNFα, 3ng/ml IL-6; (M)PE: 1ng/ml VEGF₁₆₅, 0.5ng/ml TNFα, 3ng/ml IL-6; (H)PE: 10ng/ml VEGF₁₆₅, 0.5ng/ml TNFα, 3ng/ml IL-6) circulating concentrations were applied to HUVEC. Cells exposed to the Normal cocktail were unchanged in number of ATP-stimulated Ca²⁺ bursts, and 30 minutes pretreatment with the 10uM CLA mix showed a slight reduction in Ca²⁺ bursts. Cells exposed to (L)PE and (M)PE cocktails showed modest levels of ATP-stimulated Ca²⁺ burst inhibition which were unchanged by CLA pretreatment. Exposure to the (H)PE cocktail gave severe Ca²⁺ burst inhibition, which was partially rescued using the 10uM 1:1 CLA mix. Data are presented as mean Ca²⁺ burst numbers for those cells that showed 3+ bursts on initial ATP treatment ± SEM on a minimum of 75 cells from at least 4 separate dishes. Significance by Rank-sum test is indicated as p<0.05 for * Control vs Treatment (w/ or w/o CLA); + Significant improvement after CLA pretreatment vs Treatment; # Significant exacerbation of Ca²⁺ burst inhibition by CLA pretreatment vs Treatment. Typical physiological range is shown with shaded region as defined in (Boeldt et al., 2017,Bird et al., 2013), adjusted to HUVEC.