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. 2003 Oct 27;163(2):231–236. doi: 10.1083/jcb.200304079

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Copyright © 2003, The Rockefeller University Press

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Figure 3. — Clathrin controls μ2 phosphorylation in DKO-R cells. (a) Phosphorylated μ2 is largely associated with the assembled fraction of clathrin. DKO-R cells were homogenized, and membrane (A) and cytosolic (U) fractions were prepared to separate assembled and unassembled pools of clathrin. The fractions were subjected to SDS-PAGE, and after transfer to nitrocellulose were immunoblotted with antibodies to clathrin, μ2, and phospho-Thr156 μ2. (b) Phosphorylated μ2 is substantially reduced in clathrin-depleted DKO-R cells. Exponentially growing chicken DKO-R cells treated for 96 h without (lane 1) or with (lane 2) 0.1 μg/ml doxycycline were lysed and subjected to immunoblotting with antibodies to clathrin, μ2, and phospho-Thr156 μ2.