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. 2015 Jan 8;290(10):6130–6140. doi: 10.1074/jbc.M114.616680

FIGURE 3.

FIGURE 3.

ClipR-59 preferentially interacts with Elmo2 and Elmo3. A, GST pulldown assay to examine interaction between GST-ClipR-59 and Elmo family proteins. GST-ClipR-59 or GST prepared from 239T cells was incubated with cell lysates from COS-7 cells that were transiently co-transfected with Myc-tagged Elmo expression vectors. The proteins associated with GST beads were analyzed in Western blot with anti-Myc antibody (top). The middle panel shows the levels of Myc-Elmo in TCL. The bottom panels show the levels of GST fusion proteins associated with GST beads. B, GST pulldown assay to examine the interaction between ClipR-59 and GST-Elmo proteins. Total cell lysates from COS-7 cells that were transiently co-transfected with FLAG-ClipR-59 expression vectors plus either pEBG (GST vector) or pEBG-Elmo (lanes 1–3) were subjected to a GST pulldown assay. The GST beads were analyzed in Western blot with anti-FLAG antibody (top). The middle panel shows the levels of FLAG-ClipR-59 in TCL. The bottom panels show the levels of GST fusion proteins associated with GST beads. C, similar to B except FLAG-tagged Dock180 was used. These experiments were repeated 3–4 times with similar results. IB, immunoblot.