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. 2007 May;18(5):1683–1692. doi: 10.1091/mbc.E06-09-0833

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© 2007 by The American Society for Cell Biology

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Figure 5. — Spartin binds to ubiquitin. (A) HeLa total cell lysates (TCL) were incubated with ubiquitin-agarose (Ub-beads) or protein A-agarose (Control), and aliquots from total cell lysate and bound proteins were analyzed by immunoblotting for the indicated proteins. Molecular sizes: Eps15, ∼145 kDa; STAM1, ∼62 kDa; tubulin, ∼52 kDa. (B) Lysates from HeLa cells transfected with Myc-spartin (Myc-spr) were incubated with GST, GST-Ub, or GST-Ub^I44A bound to glutathione-Sepharose. Total cell lysates (TCL) and bound proteins were analyzed by immunoblotting (WB) using an anti-Myc antibody. A Coomassie-stained gel (bottom panel) demonstrates that similar amounts of fusion proteins were used for the pulldowns.