Figure 1.

Effects of knockdown of BIG2 and/or BIG1, or AP-1 on the localization of TGN and recycling endosomal proteins. HeLa cells (a–j and p–t) or HeLa cells stably expressing CD4-furin chimera (k-o; see Materials and Methods) were mock-treated (siRNAs for LacZ; a, f, k, and p) or treated with a pool of siRNAs for BIG1 (b, g, l, and q), BIG2 (c, h, m, and r), BIG1+BIG2 (d, i, n and s), or the AP-1 μ1A subunit (e, j, o, and t), as described in Materials and Methods. The cells were then stained with antibody against TfnR (a–e), the AP-1 γ subunit (f–j), CD4 (k–o), or TGN46 (p–t). In panel s, asterisks indicate cells with reduced staining intensity for TGN46 and a plus sign indicates a cell with unaffected TGN46 staining.