Fig. 2

nsSNPs in ADGRD1 influence signaling activity in compartment-depending manner. a N-terminal nsSNPs of ADGRD1 have different influences on signaling activity and expression. The positions of nsSNPs within the N-terminal fragment (NTF) are indexed through the pictogram under the graph. b nsSNPs in 7TM core and C terminus influence cAMP response element (CRE)-signaling activity. The schema under the graph indicates the position of signaling activity relevant nsSNPs: S667L in intracellular loop (ICL) 2, V764M in TM6 (VI), N795K in TM7 (VII) and A816T and T827M in C terminus. c Constitutive activity of P2Y₁₂-CTF-mutant is lost after insertion of N795K. (A-C) Graphs show the percentage of wildtype (wt) after normalization to mock control of functional relevant nsSNPs in CRE-SeAP activity (EV: 568,311 ± 59,100 counts; wt: 786,125 ± 85,787 counts; n = 4), cell surface expression (EV: 0.02 ± 0.03 OD_{492 nm}; wt: 0.70 ± 0.11 OD_{492 nm}; n = 9) and whole cell expression (EV: 0.08 ± 0.07 OD_492nm; wt: 0.97 ± 0.16 OD_492nm; n = 7). To compare differences of receptor mutants to wt basal activity or expression levels an unpaired two-tailed t-test was performed with * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. d Activation analysis of nsSNPs upon stimulation with the tethered agonist-derived peptide p13. After normalization to mock control (EV: 872 ± 200 counts; n = 7) data are shown as percentage of wildtype stimulated with p13 (wt + p13: 5.3 ± 1.3 x-fold over EV; n = 7). To analyze significance of receptor mutant activation after p13 stimulation a two-way ANOVA with Bonferroni as post-test was used: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. a-d Data are given as means ± SD