Inducible expression of α2B-adrenergic receptor

The Tet-On 3G Tetracycline Inducible Gene Expression System (Clontech Laboratories, Inc.) was utilized to generate stable cell lines inducibly expressing HA-α_2B-AR in HEK293 cells as described previously^{17 (link),35}. Briefly, HA-α_2B-AR was cloned into the pTRE3G-TRES vector at the BglII and ClaI restriction sites and co-transfected with the PLKO.1 vector in HEK293 cells using Lipofectamine 2000. Inducible α_2B-AR expression was verified by intact cell ligand binding assays, immunoblotting and confocal microscopy³⁵. The current study uses a cell line that expresses 8.5 × 10⁵ receptors per cell.

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Zhang M., Xu X., Li C., Huang W., Xu N, & Wu G. (2019). A Naturally Occurring Splice Variant of GGA1 Inhibits the Anterograde Post-Golgi Traffic of α2B-Adrenergic Receptor. Scientific Reports, 9, 10378.

Publication 2019

Tet-On 3G Tetracycline Inducible Gene Expression System

Assays Cell Cell line Cell receptors Gene expression Hek293 cells Ligand Lipofectamine 2000 Tetracycline Vector

Corresponding Organization :

Other organizations : Guangzhou University of Chinese Medicine, Augusta University

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Variable analysis

independent variables

Inducible expression of HA-α2B-AR in HEK293 cells

dependent variables

α2B-AR expression verified by intact cell ligand binding assays, immunoblotting and confocal microscopy

control variables

Cell line that expresses 8.5 × 10^5 receptors per cell

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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