Fig. 4
From: Analysis of CCR2 splice variant expression patterns and functional properties
Membrane localization of CCR2 variants. A HEK293 cells were transfected with plasmids containing FLAG or HA-tagged CCR2 variant sequences and lysed with radioimmunoprecipitation assay (RIPA) buffer. The clarified cell extracts were subjected to western blotting using the appropriate antibodies. B HEK293 cells on coverslips were transfected with GFP-tagged forms of CCR2 variants and fixed with 4% paraformaldehyde, and the GFP signals were observed under a confocal microscope. C Different quantities of plasmids containing the N-terminal SmBiT-tagged chemokine receptor sequences were transfected in HEK293 cells. HiBiT activities were measured by a luminometer at a single time point. CCR2C1: the chimeric form of CCR2 in which the C-terminal region was replaced with the C-terminal region from CCR1. D Helix 8 in the C-terminal regions of CCR2A, CCR2B, and CCR1. Yellow color designates Helix 8. The red color indicates basic amino acids. E Cells co-expressing SmBiT-β-arrestin 1 with CCR2B-LgBiT or CCR2C1-LgBiT were treated with monocyte chemoattractant protein-1 (MCP-1), and luciferase activity was measured by a luminometer. F The basal luciferase activity was measured in cells expressing both the SmBiT and LgBiT forms of the receptors