Figure 2.

ORP1L and RILP colocalize and are part of a physical complex. (A) ORP1L and RILP colocalize on juxtanuclear late endocytic clusters. HeLa cells were transfected with GFP–RILP or GFP–ΔN-RILP, together with Xpress-tagged ORP1L or Xpress-tagged ORP1L–ANK domain (red), as indicated in the images. ANK and ORP1L colocalize with RILP or ΔN-RILP (right, merge). Bars, 10 μm. (B) ORP1L co-isolates with expressed RILP. HeLa cells were transfected with Xpress-tagged RILP and immunoprecipitated with anti-Xpress antibody (α-Xpress) or irrelevant mouse IgG (MIgG). The isolates were Western blotted with anti-ORP1L antibodies (WB: α-ORP1L). (bottom) Corresponding lanes probed with anti-Xpress antibody (WB: α-Xpress). (C) Coimmunoprecipitation of endogenous RILP and ORP1L. HeLa cell lysates were immunoprecipitated with rabbit anti-RILP serum (α-RILP) or irrelevant rabbit IgG (RIgG). The isolates were Western blotted with anti-ORP1L antibodies (WB: α-ORP1L).