Figure 4: Inhibition of ATP-stimulated Ca²⁺ bursts is partially rescued by a 10uM 1:1 mix of c9,t11 and t10,c12 CLA isomers for VEGF₁₆₅, TNFα, and bFGF.

Confluent HUVEC were pre-exposed to a 1:1 mix of either 50uM c9,t11 CLA and 50uM t10,c12 CLA or 10uM c9,t11 CLA and 10uM t10,c12 CLA for 30 minutes before growth factor or cytokine treatment and subsequent 100uM ATP treatment. ATP-stimulated Ca²⁺ bursts remained significantly reduced compared to control for all CLA-exposed treatments. The 50uM CLA mix failed to improve ATP-stimulated Ca²⁺ bursts for any growth factor or cytokine and exacerbated the Ca²⁺ burst inhibition for EGF (panel C), IL-6 (panel F), and IL-8 (panel G). The 10uM CLA mix significantly improved ATP-stimulated Ca²⁺ bursts for VEGF₁₆₅ (panel A), bFGF (panel B), and TNFα (panel D). The 10uM CLA mix exacerbated Ca²⁺ burst loss for IL-1β (panel E) and IL-8 (panel G). Data is 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.