Fig. 6: Chemokine receptor CXCR7 interacts with SHLP2.

a Schematic for PathHunter β-Arrestin assay. b Activity plot showing the results of PathHunter β-Arrestin GPCR screening. Note that CXCR7 exhibited the highest activity by SHLP2. c Dose-dependent β-Arrestin recruitment to CXCR7 by SHLP2. The concentration of SHLP2 inducing half-maximal response (EC₅₀) was calculated by least-square fitting to a four-parameter logistic curve. d Pulldown analysis of streptavidin-SHLP2-biotin showing the interaction between SHLP2 and CXCR7. Data shown are representative of five independent experiments with similar results. e Schematic (upper panel) and the plasmid DNA (lower panel) of the NanoBiT complementation assay to monitor SHLP2-induced β-Arrestin2 recruitment to CXCR7. f Results of β-Arrestin2 recruitment to CXCR7 by SHLP2 or CXCL12 (n = 4). g Western blot result of time-dependent ERK1/2 phosphorylation by SHLP2 with or without CXCR7 overexpression. Data shown are representative of five independent experiments with similar results. h Representative spike of POMC neuron showing the depolarization by SHLP2 is blocked by the pretreatment of the MAP Kinase blocker PD98059. i Summary graph of SHLP2 effect on the resting membrane potential (RMP) of POMC neurons under treatment of the MAP Kinase blocker (n = 10 from 4 animals; ns: not significant, paired t test). j Representative depolarizing response of POMC neurons by SHLP2 under treatment of the PI3K inhibitor wortmannin. k Summary graph showing the changes in resting membrane potential (∆RMP) by SHLP2 treatment with or without MAP Kinase or PI3K inhibitor (n = 27, 10, 5, respectively). n indicates the number of biologically independent cells examined. Data are presented as the mean ± SD. PK, enzyme donor fragment ProLink; β-Arr2, β-Arrestin2; β-gal, β-galactosidase; Nluc Nano Luciferase, SmBiT Small fragment, LgBiT Large fragment, UBC pro UBC promotor. Source data are provided as a Source Data file.