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. 2007 Feb 12;176(4):459–471. doi: 10.1083/jcb.200606077

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Copyright © 2007, The Rockefeller University Press

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Figure 6. — RILP interacts with the C-terminal 25-kD fragment of p150^Glued. (A) HeLa cells were transfected with mRFP–RILP or mRFP-tagged ΔN-RILP, along with GFP-tagged ΔC and C25 fragments of p150^Glued. (B) HeLa cells were transfected with GFP–p150^Glued C25 or GFP–p50^dynamitin, fixed, and stained with anti-CD63 antibodies. (C) Purified GTPγS-loaded Rab7Q67L coupled to GST-beads was incubated with combinations of purified RILP, ΔN-RILP, ORP1L, or p150^Glued C25 (aa 1,049–1,278), as indicated above the gels. Proteins bound in a GTP-dependent manner were eluted with EDTA and the C-terminal MBP–p150^Glued fragments (C25) were captured on amylose resin. The input and resin-bound proteins were Western blotted with anti-RILP, -GST, or -MBP antibodies. Bars, 10 μm.