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. 2000 Oct;11(10):3397–3410. doi: 10.1091/mbc.11.10.3397

Figure 4.

Figure 4

Constitutive phosphorylation of 70z-Cbl when expressed in MDCK cells. (A) Scheme of 70z-Cbl and its mutant variants. PTB/SH2, phosphotyrosine-binding SH2 domain; RF, RING finger domain; Pro, proline-rich domain. (B) 70z-Cbl and its variants were immunoprecipitated from MDCK cell populations, separated on an 8% SDS-PAGE gel, and immunoblotted with anti-HA (upper panel) or anti-phosphotyrosine (lower panel) antibodies. (C) Lysates from MDCK cell populations were incubated with the amino-terminal SH2 domain of p85 (GST-N-SH2 p85; upper panel) or the amino- and carboxyl-terminal SH3 domains of Grb2 (GST-N+C-SH3 Grb2; lower panel). Proteins were separated on an 8% SDS-PAGE gel, transferred to nitrocellulose, and immunoblotted with anti-HA antibody. (D) MDCK cell populations were immunoprecipitated with the indicated antibodies, resolved by SDS-PAGE, transferred to nitrocellulose, and immunoblotted with anti-HA antibody.