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. Author manuscript; available in PMC: 2010 Sep 7.
Published in final edited form as: Nat Immunol. 2008 Oct 19;9(12):1356–1363. doi: 10.1038/ni.1670

Figure 6.

Figure 6

Nedd4 binds to and promotes ubiquitination of Cbl-b. (a, b) Primary CD4+T cells were isolated from Nedd4+/+ and Nedd4−/−fetal liver chimeras. Cells were cultured for 24 h with or without stimulation as noted. Whole cell lysates were (a) immunoblotted with antibodies specific for Cbl-b, Nedd4 or actin (loading control), or (b) immunoprecipitated with anti-Cbl-b and immunoblotted with anti-ubiquitin. (c) Primary CD4+ T cells were isolated from Nedd4+/+ and Nedd4−/−fetal liver chimeras. Cells were cultured for 24 h in the absence or presence of plate-bound anti-CD3 and anti-CD28. Lysates were immunoprecipitated with anti-Nedd4 where indicated, and immunoblotted for Nedd4, Cbl-b and actin (loading control). WCL, whole cell lysate. These data are representative of 2–3 independent experiments.