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Beads were then washed four times with wash buffer (50 mM TrisHCl pH 7.4, 150 mM NaCl, 0.1% Triton X-100) and then twice with wash buffer lacking Triton X-100

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The level and transcription of RACK1, the receptor for activated C kinase, a regulator of G1/S progression, which does not contain a consensus HECT or RING-type domain characteristic of typical E3 ubiquitin ligases, 57 but binds to and promotes the ubiquitination of p63 and p73, was not altered in any cancer or normal cell line (Figures 7 and 8) in response to p28