Figure 3

MCOLN1 RNAi clones. A) Northern blot done on 15 μg of total RNA isolated from RAW264.7, LS9, or LS10 cells. The same filter was probed for MCOLN1, stripped, and re-probed for GAPDH, stripped again and re-probed for MCOLN2. B) Confocal images of RAW264.7, LS9, and LS10 cells stained with Acridine Orange (AO). Bar is 6 μm. C) Quantitation of the intensity of staining of AO-stained compartments and of dextran-Oregon Green (OG)-stained compartments. Bars represent standard deviations. D) Confocal images of RAW264.7, LS9, and LS10 cells whose terminal compartments were pre-loaded with Dextran-Cascade Blue (DEX-CB, red). BSA-Bodipy LacCer (B-LACCER, green) was added for 30 minutes and chased for the indicated times before fixation. Arrows indicate co-localization of the two markers. Bar is 5 μm.