Extended Data Fig. 2: Identification and optimization of nanobodies as PAMs of ADGRG2.
From: Development of an allosteric adhesion GPCR nanobody with therapeutic potential
a. Representative dose-response curves from three independent experiments of two ADGRG2 FlAsH-BRET sensors in response to Nb23, Nb32 or Nb6 stimulation (n = 3) are shown. The EC50 values are the mean ± SEM from three independent experiments performed in triplicate (n = 3). b. Representative saturation binding curves from three independent experiments of Nb23-FITC, Nb32-FITC, Nb23-bi-FITC, Nb32-bi-FITC toward Nluc–ADGRG2 (n = 3) are shown. FITC is conjugated at the C terminus of the nanobodies. c, Representative dose–response curves from three independent experiments of DHEA-induced cAMP accumulation in mADGRG2-overexpressing HEK293 cells treated with or without 1 μM nanobodies, as determined by the GloSensor assay (n = 3) are shown. The curve representing DHEA-induced ADGRG2 activation in right panel is a replica of the corresponding curve in left panel, depicted as a dotted line to facilitate comparison and enhance clarity in the evaluation. The EC50 values are presented as the mean ± SEM from three independent experiments performed in triplicate (n = 3). d. Representative dose-response curves from three independent experiments of DHEA-induced InsP3 (IP3) production in mADGRG2-overexpressing HEK293 cells treated with or without 1 μM nanobodies (n = 3) are shown.
