Fig. 5.

Effects of Gβγ inhibition on its associations with downstream effectors. A: human coronary artery endothelial cells (HCAECs) were pretreated with DMSO or the Gβγ-subunit inhibitor gallein (30 μM) for 30 min before being subjected to flow for 15 s. In situ proximity ligation assay (PLA) was performed using antibodies against Gβ₁ and the p101 subunit of phosphoinositide 3-kinase (PI3K). Representative images are depicted with the bar graph showing quantification of 3 independent experiments as PLA signal (green dots) per cell (blue nuclei) relative to the DMSO static condition. Error bars indicate SE. ***P < 0.001. B: HCAECs were pretreated with DMSO or gallein (30 μM) for 30 min before 15-s flow. In situ PLA was performed using antibodies against Gβ₁ and Piezo1. Representative images are depicted with the bar graph showing quantification of 4 independent experiments as PLA signal (green dots) per cell (blue nuclei) relative to the DMSO static condition. Error bars indicate SE. ***P < 0.001. C: HCAECs were pretreated with DMSO or gallein (30 μM) for 30 min before stimulation with ${\text{AlF}}_4^{-}$ (30 μM AlCl₃/10 mM NaF) for 1 min. In situ PLA was performed using antibodies against Gβ1 and Piezo1. Bar graph shows quantification of 3 independent experiments as PLA signal per cell relative to the DMSO static condition, with error bars indicating SE. ***P < 0.001.