Fig. 5: Mapping the interaction region between GABAB receptor and integrin β3.
From: GABA-independent activation of GABAB receptor by mechanical forces
a 3D model of the VFT of GB1 and GB2. N-glycosylation mutations (M1-7) are highlighted in LB2 of GB1 VFT. b Detailed presentation of the mutants of M1-7 and 5 A. c Schematic representation of GABAB-M1-7 and GABAB-5A. d IP1 production in HEK293 cells transfected with GABAB receptor WT, or GABAB receptor mutants M1-7 along with Gqi9 under suspension or adhesion conditions. Data are present as mean ± s.e.m. from at least three biologically independent experiments (from left to right: n = 6, 6, 6, 5, 6, 5, 5, 4, 5) each performed in triplicates and analyzed using paired t test (two-tailed) to determine significance. **P < 0.01, *P < 0.05, not significant (ns) > 0.05. e Intracellular calcium release induced by different dose of GABA in HEK293 cells transfected with GABAB receptor WT and Gqi9, or GABAB-5A and Gqi9. Data are presented as mean ± s.e.m. from three biologically independent experiments each performed in triplicates. f IP1 production in HEK293 cells transfected with GABAB receptor WT and Gqi9, or GABAB-5A and Gqi9 under suspension or adhesion conditions. Data are mean ± s.e.m. from four biologically independent experiments each performed in triplicates and analyzed using unpaired t test (two-tailed) to determine significance. *P < 0.05, not significant (ns) > 0.05. g Real-time recording of intracellular Ca2+ release in HEK293 cells expressing GABAB-5A and Gqi9. After recording the basal state of Ca2+ release for 50 seconds, cells were subjected to shear stress for 100 seconds. Shear stress was then halted for 150 seconds, after which baclofen was added and the Ca2+ signal was measured for another 200 seconds. Data are present as mean ± s.e.m. from 111 cells recorded.
