Cells were lysed in RIPA buffer supplemented with a protease inhibitor cocktail (Calbiochem) as described [17 (link)]. Equal amounts of protein were separated by SDS-PAGE, transferred to Immobilon-FL polyvinylidene fluoride (PVDF) transfer membrane (Millipore), and immunostained. Immunoblots were visualized using an Odyssey infrared scanner (LI-COR).
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