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. 2003 Jun 2;197(11):1511–1524. doi: 10.1084/jem.20021686

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Copyright © 2003, The Rockefeller University Press

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Figure 3. — The activation-induced ubiquitination of Syk is reduced in Cbl-b^−/− B cells. Splenic B cells from Cbl-b^−/− and Cbl-b^+/+ mice were treated as in Fig 2, A–C, to cross-link the BCR and warmed to 37°C for increasing lengths of time. The cells were lysed, and Syk was immunoprecipitated and analyzed by SDS-PAGE and immunoblotting. Duplicate immunoblots were probed for either ubiquitin (anti-Ub) or phosphotyrosine (Anti-PTyr). The phosphotyrosine blot was stripped and reprobed for Syk (anti-Syk). The results for the 2-min time point are shown at which point ubiquitination of Syk was maximal. The result shown is representative of three independent experiments.