Figure 4.

Down-regulation of p110α and p110δ inhibits lysoPC-induced increase in [Ca²⁺]_i. Bovine aortic ECs were transiently transfected with control siRNA (NsiRNA) (40 nmol/L), p110α siRNA (20 nmol/L), p110β siRNA (30 nmol/L), p110δ siRNA (20 nmol/L), or p110γ siRNA (30 nmol/L) for 24 h. ECs were loaded with the FITC-conjugated fluorophore Calbryte 520 AM dye. The EC were suspended and loaded into the sort chamber of a BD FACSMelody cell sorter maintained at 37°C. After adjusting the baseline, lysoPC (12.5 μmol/L) was added. A–D: Using the kinetic reading mode at Ex/Em 490/525 nm, relative changes in [Ca²⁺]_i after transfection with NsiRNA (A), p110α (B), p110β (C), p110δ (D), or p110γ (E) were determined (Representative images of n = 3 independent biologic samples). F: Fold increase of [Ca²⁺]_i measured by difference in mean [Ca²⁺]_i at baseline and after addition of lysoPC (12.5 μmol/L) presented as a dot whisker plot (n = 3 independent biological samples; *P = 0.02 compared with NsiRNA, †P = 0.01 compared with NsiRNA). Statistical analysis performed using One-Way ANOVA with Dunnett’s Multiple Comparison Test against NsiRNA.