Figure 1. Tollip interactions.

A) GST-Tollip interaction with ³⁵S-methionine labelled proteins isolated by the yeast two hybrid technique. Lane 1 contains 1/10 input of the ³⁵S-methionine protein used for each interaction; lanes 2 and 3 contain the elution product from incubation of the ³⁵S-proteins with the GST-Tollip and GST-protein alone, respectively. ³⁵S-methionine labelled proteins were visualized by autoradiography. Western blot analysis of 293T cells transfected with Ubc9 and HA-Tollip (B), Flag-ARIP3 and HA-Tollip (D), HA-Cystatin B and Ubc9 (C), HA-Cystatin B and Flag-ARIP3 (E). HA-Cystatin B is a ubiquitous protein with antiprotease function, unrelated to Tollip, nor to the inflammatory pathway [28]. Lane 1 contains the protein extract; lane 2 contains the proteins immunoprecipitated with anti-HA abs. In this and in the following figures, “pe.” refers to the protein extract and “ip.” to the immunoprecipitated protein. Staining carried out as indicated under the figures.