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. Author manuscript; available in PMC: 2010 Jun 15.
Published in final edited form as: Gene. 2009 Mar 28;439(1-2):87–94. doi: 10.1016/j.gene.2009.03.014

Figure 1. In vivo and in vitro interactions between Cux1 and Grg4.

Figure 1

A: Kidney lysates from 3-day-old wild type (WT) and Cux1 transgenic (TG) mice were immunoprecipitated with rabbit polyclonal anti-Cux1 (top panel) or rabbit polyclonal anti-Grg4 (bottom panel) as indicated. The immunoprecipitates were then blotted for Grg4 (top panel) or Cux1 (bottom panel) as indicated. Grg4 can be seen in WT and TG kidneys, after immunoprecipitation with Cux1 (right lanes, top panel). Cux1 can be seen in WT and TG kidneys, after immunoprecipitation with Grg4 (right lanes, bottom panel). No bands are observed when protein G beads alone are precipitated (no Ab). B: Bacterially expressed Cux1 protein was incubated with GST- Grg4 fusion protein immobilized on Glutathione agarose beads. Following washing, GST beads alone with Cux1, wash #3, and GST-Grg4 immobilized beads with Cux1 were transferred to a membrane and blotted with antibody directed against Cux1 protein. Only GST-Grg4 with Cux1 protein and the Cux1 input showed a positive band, indicating a direct physical interaction between Cux1 and Grg4.Lane 1 shows 50% of the total purified recombinant Cux1 protein used for the assay.