Extended Data Fig. 2: Immunoblotting of Best2 and GS in HEK293 cells.
From: Bestrophin-2 and glutamine synthetase form a complex for glutamate release
(a) Indicated Best2-YFP-His and GS-CFP-Myc constructs were co-expressed and detected by an anti-GFP antibody in input (left) and co-immunoprecipitation (right) samples. The last lane on the IP gel is performed in the presence of 20 mM glutamate. (b) The overall, cytosolic and membrane-bound protein levels of WT hBest2-YFP-His and hGS-CFP-Myc upon individual- or co-expression were detected by an anti-GFP antibody. (c) Exogenous hGS-CFP-Myc and endogenous hGS in HEK293 cells were detected by an anti-GS antibody. (d) The cytosolic and membrane-bound levels of indicated hBest2-YFP-His and hGS-CFP-Myc constructs upon co-expression were detected by an anti-GFP antibody. (e) hBest2-YFP-His and hGS-CFP-Myc were individually co-transfected with hBest2 siRNA and hGS siRNA, respectively, or a scramble siRNA, and detected with an anti-GFP antibody (left). β-Actin (right) served as the loading control on the same blot. Each experiment was biologically replicated three times with similar results.
