Figure 1: Nedd4 is an IRS-2-associated protein.
From: Nedd4-induced monoubiquitination of IRS-2 enhances IGF signalling and mitogenic activity
(a,b) Determination of the binding regions. HEK293T cells overexpressing the indicated FLAG-tagged IRS-2 deletion mutants (a) or myc-IRS-2 (b) were serum starved and cell lysates were subjected to pull-down analysis using GST-Nedd4 or its deletion mutants shown in b. The purity of GST-tagged proteins was validated by SDS–PAGE and CBB stain. Input, ∼1/100 volume of HEK293T lysates. *GST-Nedd4 Δ227–551 was not successfully expressed. (c,d) Effects of Nedd4 HECT domain (c) and various Nedd4 mutants (d) on the IRS-2–Nedd4 C2 domain interaction. HEK293 cells overexpressing indicated proteins were serum starved and cell lysates were subjected to immunoprecipitation and immunoblotting using the indicated antibodies.
