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. 2016 Jul 14;6:29205. doi: 10.1038/srep29205

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Copyright © 2016, Macmillan Publishers Limited

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(A) CFP-pIEx-His (blue) and YFP-pIEx-His (yellow) plasmids were co-transfected in cells as controls. (B) ErGPCR2-CFP-pIEx-His and GRK2-YFP -pIEx-His plasmids were co-transfected in the cells. Cells were treated with DMSO or 20E (1 μM) for 5 min. Merge is the overlapped yellow and blue. The yellow bar denotes 20 μm. (C) Ap-Fret detected the interaction between ErGPCR-2 and GRK2. ErGPCR-2-CFP and GRK2-YFP plasmids were co-tranfected in cells. The overexpression of ErGPCR-2-CFP was the negative control. Cells were incubated with DMSO or 20E (1 μM for 5 min). Ap-Fret efficiency (%) = (Ipost − Ipre) × 100/Ipost, the Ap-Fret efficiency of ErGPCR-2-CFP overexpression as the negative control. Asterisks indicate significant differences between the groups (p < 0.05) by the student’s t-test based on three independent experiments.