Western Blot Detection and Visualization

Western blot was performed as described previously52 (link)53 (link)54 (link). In brief, equal amounts of cell lysates were resolved by SDS-PAGE and then transferred to polyvinylidene fluoride or nitrocellulose membranes. Membranes were blocked and incubated overnight with primary antibodies at 4 °C. The membranes were incubated with appropriate HRP-conjugated secondary antibodies at room temperature for 1 h, and the blots were visualized with PlusECL (KeyGEN BioTECH) according to the manufacturer’s protocol. Alternatively, blots were detected with IRDye 800 CW conjugated anti-rabbit IgG or IRDye 680 CW conjugated anti-mouse IgG secondary antibodies (LI-COR Biosciences, Lincoln, NE), and visualized using Odyssey infrared imaging system (LI-COR Biosciences).