FIG 2.

Molecular requirements for RIDα-induced cholesterol trafficking. (A) Equal protein aliquots immunoblotted with antibodies to specific siRNA targets listed in the figure or actin following treatment with noncoding (NC) or target-specific siRNAs. (B) CE quantification (in nanograms of cholesterol per milligram of protein) in shNPC1-RIDα cells treated with target-specific siRNAs. Data are normalized to NC siRNA-treated cells (means ± s.e.m.; n = 3; **, P < 0.005). (C and D) Representative confocal images of all hepatocyte-derived cells following treatment with NC siRNA (C) and shNPC1-RIDα cells treated with the target-specific siRNAs listed in the figure (D). Cells were stained with Bodipy 493/503 (green) and DAPI (blue) and FLAG epitope antibody to detect FLAG-RIDα (red). (E and F) Representative confocal images of all hepatocyte-derived cells following treatment with NC siRNA (E) or of shNPC1-RIDα cells treated with target-specific siRNAs. Cells were stained with LAMP1 antibody (red), filipin (blue), and FLAG-RIDα antibody (green). Boxed areas indicate regions are magnified at ×2 to visualize individual and merged channels to the right of each image. (C to F) All size bars, 5 μm.