| Autor: |
Nguyen, Huong Thi, Arfaxad Reyes-Alcaraz, Yong, Hyo Jeong, Nguyen, Lan Phuong, Park, Hee-Kyung, Inoue, Asuka, Lee, Cheol Soon, Seong, Jae Young, Hwang, Jong-Ik |
| Rok vydání: |
2020 |
| DOI: |
10.6084/m9.figshare.13279119.v1 |
| Popis: |
Additional File 1: Fig. S1. Ligand-stimulated real-time luciferase activities in HEK293 cells expressing different combinations of NanoBit constructs ofCXCR7 and β-arrestin2. Fig. S2. Ligand-stimulated real-time luciferase activities in HEK293 cells expressing different combinations of NanoBit constructs of β-arrestin2 with CXCR4 or CXCR3. Fig. S3. Receptor internalization assay using NanoBit constructs. (a) Cells expressing receptor-LgBiT and SmBiT-FYVE domain of EEA1 were treated with SDF-1α and the luciferase activities were measured in real-time. (b) Cells expressing receptor-SmBiT and LgBiT-CAAX sequence were used in the NanoBit assay. Fig. S4. Optimization of NanoBit construct combinations of Gb1 and GRKs. a and b showed luciferase activities in cells expressing different combinations of NanoBiT constructs. Fig. S5. Optimization of NanoBit construct combinations of receptor and GRK2. Fig. S6. Generation of cells lacking receptors using CRISPR system. (a) Genomic DNA PCR products from cells established with CRISPRCas9were cloned and sequenced. Red color designates guide RNA target regions. (b) RT-PCR products of either CXCR4 or CXCR7 were compared in wild-type and receptor KO HeLa cell clones. β-actin products were used as the control. (c) Membrane expression of exogenous receptors was not affected by deletion of CXCR4 or CXCR7. HiBiT constructs of the receptors were expressed in wild-type and receptor KO of HEK293 and HeLa cells, and the cells were applied to the HiBiT assay. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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