Anti kv11

HL-1 cells were lysed using RIPA buffer (Thermo Fisher, Waltham, MA) containing protease inhibitor cocktail (Roche, South San Francisco, CA). Equal amounts of protein (10–20 μg) were loaded into wells of 7% or 3–8% tris-acetate gels, subjected to SDS-PAGE, and transferred to a 0.45 μm PVDF membrane (Thermo Fisher). Western blot analyses were performed using the following primary antibodies: anti-Flag (Sigma-Aldrich), anti-Kv4.3 (Alomone, Jerusalem, Israel), anti-Kv11.1 (Alomone), anti-Nav1.5 (Alomone), anti-Cav1.2 (Alomone), and anti-GAPDH (Millipore, Billerica, MA). The immunodensity of bands corresponding to targeted proteins were quantified using NIH ImageJ software, and values were corrected for protein loading by dividing them by GAPDH immunodensity signals in the same lanes. Subsequently, corrected values obtained from Western blots loaded with protein lysate from BKα-transfected cells were normalized to corrected values obtained from Null-transfected cells corresponding to the same ion channel. According to this analysis, by definition the average normalized intensity of Null transfection is unity. For Western blots designed to detect Kv11.1, both bands of the Kv11.1 doublet revealed by anti-Kv11.1 blotting were included in calculations.