Figure 4.

Ceramide Does Not Affect PS-ASO Trafficking

(A) Representative immunofluorescence images of cells pretreated with 10 μM ceramide or untreated (control) for 6 hr, incubated with 2 μM Cy3-PS-ASOs (IONIS ID 446654) for 2 hr and stained for lipid droplets (green) and PS-ASOs (red). Nuclei were stained with DAPI (blue). Merged images are also shown. Scale bars, 2 μm. (B and C) Representative immunofluorescence images of cells pretreated with 10 μM ceramide or untreated (control) for 6 hr, incubated with 2 μM Cy3-PS-ASO for 2 hr, and stained for (B) EEA1 (green) or (C) LBPA (green) and LAMP1 (green) and PS-ASO (red). Nuclei were stained with DAPI (blue). Scale bars, 2 μm. The PS-ASO-positive early endosomes or late endosomes were counted in 30 cells, and the percentage of the PS-ASO-positive early endosomes or late endosomes was calculated relative to the total numbers of the PS-ASO-positive organelles.