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Immunoblotting was carried out as previously described [26 (link)]. Briefly, equal amounts of whole-cell extracts or equal volumes of pull-down material were separated by 12% SDS-PAGE under denaturing conditions and transferred to nitrocellulose membrane (Protran; Amersham/Merck, St. Louis, MO, USA). The nitrocellulose membranes were probed with the appropriate primary antibodies after blocking with Tris-buffered saline (TBS) containing 5% nonfat milk and 0.1% Tween-20 for 1 h at RT. Except for the neutravidin-HRP conjugate diluted to a final concentration of 1:2000, all primary antibodies were diluted in blocking buffer at 1:1000. Secondary HRP-conjugated antibodies were used in a 1:2000 dilution. The immunoreactive bands were visualized with the enhanced chemiluminescence (ECL) method using the Clarity substrate (BioRad). Data obtained from at least three independent experiments are presented here.