Figure 5.

GPR35-mediated intracellular signaling.A, reduction of forskolin-induced cAMP levels elicited by imetit in histamine H₃ receptor (H₃R)-transfected cells. B, recruitment of p63RhoGEF-RlucII to the plasma membrane upon activation of thromboxane A₂ receptors (TBXA₂R) with U-46619. C, generation of DAG upon activation of TBXA₂R with U-46619. D, generation of cAMP upon activation of histamine H₂ receptors (H₂R) with histamine. E, recruitment of the receptor-binding domain of protein kinase N (PKN-RBD) to the plasma membrane upon activation of TBXA₂R with U-46619. F–H, reduction of forskolin-induced cAMP second messenger levels upon stimulation of pcDNA-, GPR35 short-, or GPR35 long-transfected cells with pamoic acid (F), kynurenic acid (G), or zaprinast (H). I–K, recruitment of p63RhoGEF-RlucII to the plasma membrane upon stimulation of pcDNA-, GPR35 short-, or GPR35 long-transfected cells with pamoic acid (I), kynurenic acid (J), or zaprinast (K). L–N, generation of DAG upon stimulation of pcDNA-, GPR35 short-, or GPR35 long-transfected cells with pamoic acid (L), kynurenic acid (M), or zaprinast (N). O–Q, generation of cAMP upon stimulation of pcDNA-, GPR35 short-, or GPR35 long-transfected cells with pamoic acid (O), kynurenic acid (P), or zaprinast (Q). R–T, PKN-RBD recruitment to the plasma membrane upon stimulation of pcDNA-, GPR35 short-, or GPR35 long-transfected cells with pamoic acid (R), kynurenic acid (S), or zaprinast (T). Data show mean ± s.e.m. of three to four independent experiments conducted in transiently transfected HEK293A cells. Vehicle-treated ΔBRET and ΔFRET values were set to 0. DAG, diacylglycerol; PKN-RBD, receptor-binding domain of protein kinase N; BRET, bioluminescence resonance energy transfer; HEK293, human embryonic kidney 293A.