Western Blotting of Recombinant Proteins

Protein preparations for Western blotting from transfected HEK293S cells and tissues are described in the Supplemental Information. Protein samples (1–100 µg, Table S6) were loaded alongside the Benchmark (Cat# 10748010, Life Technologies, Carlsbad, CA, USA), PageRuler (Cat# 26616, Thermofisher, Waltham, MA, USA) or ab116027 (Abcam, Cambridge, UK) protein ladders onto 4–12% SurePage Bis-Tris gels (GenScript, Piscataway, NJ, USA). Western blotting was performed as previously described [43 (link)]. Under some conditions, 2-(N-morpholino)ethanesulfonic acid (MES) running buffer was used to aid in the separation of lower molecular weight bands. Figure legends provide detailed information about the conditions used.