FIG. 2.

Intracellular trafficking of AONs during gymnosis. (A) Confocal images of DM500 EGFP cells after 24 h of growth in the presence of Cy3-CAG7-OMePS or Cy3-DMD23-OMePS. Lamp-1, a lysosomal marker protein, was visualized by immunochemistry and co-localized with some of the AON-positive vesicles. Scale bars indicate 25 μm. (B) Uptake of Cy3-DMD23-OMePS and Cy3-CAG7-OMePS (500 nM) in the presence of chloroquine or dynasore after 24 h of gymnosis. Merged images are a composite of Cy3 and EGFP channels. Scale bars indicate 25 μm. (C) Quantification of expanded DMPK RNA expression after 24 h of gymnosis incubation with Cy3-CAG7-OMePS in combination with chloroquine or dynasore. DMPK mRNA levels after Cy3-CAG7-OMePS and control AON treatments were compared by unpaired t-tests. CAG7-OMePS activity in the presence of chloroquine compared with vehicle was analyzed by one-way ANOVA followed by a Bonferroni's Multiple-Comparison Test (*p < 0.05, **p < 0.01).