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. Author manuscript; available in PMC: 2009 Oct 10.
Published in final edited form as: J Mol Biol. 2008 Jul 25;382(3):638–651. doi: 10.1016/j.jmb.2008.07.045

Fig. 3. Trak1 and Hrs associate in vivo and in vitro.

Fig. 3

(a) Co-immunoprecipitation of Trak1 with Hrs in transfected HeLa cells. HeLa cells were co-transfected with pGFP-Trak1 in combination with pHA-Hrs or pHA vector. Lysates were immunoprecipitated with anti-HA antibody, followed by immunoblotting with anti-GFP and anti-HA antibodies. The asterisk indicates a non-specific band. (b) Co-immunoprecipitation of endogenous Hrs with Trak1 in HeLa cells. HeLa cell lysates were immunoprecipitated with anti-Trak1 antibody or pre-immune serum, followed by immunoblotting for Hrs and Trak1. (c) In vitro association between Hrs and Trak1. Soluble His-tagged Hrs was incubated with equal amounts of immobilized GST or GST-Trak1 fusion proteins. Bound His-Hrs and immobilized GST-fusion proteins were detected by immunoblotting with anti-Hrs, anti-Trak1 and anti-GST antibodies.