Volume 8 issue 7 July 2007
A comprehensive model for the cellular uptake of cationic cell-penetrating peptides
F Duchardt, M Fotin-Mleczek, H Schwarz, R Fischer and R Brock

Supplemental Figure 1. The effect of inhibitors of endocytosis on the internalization of tracer molecules. HeLa cells were incubated for 30 min at 37°C with the indicated inhibitors of endocytosis (CPZ 10 µg/ml, MβCD 5 µM, EIPA 100 mM) or remained untreated (control groups).Then the medium was removed and cells were incubated for further 30 min with either a transferrin Alexa Fluor 633 conjugate (10 µg/ml), a dextran Alexa Fluor 647 conjugate (10 µM) or Cholera Toxin Subunit B Alexa Fluor 555 (10 µg/ml) in the presence or absence of the corresponding inhibitor. The scale bars represent 20 µm.

Figure 1 (jpg)

Supplemental Figure 2. The effect of endocytic inhibitors on cell viability. HeLa cells were incubated with inhibitors of endocytosis, either alone or in combination, at the indicated concentrations for 2 hours at 37°C. Cell viability was determined using crystal violet staining. The frames indicate the concentrations of inhibitors applied in the experiments.

Figure 2 (jpg)

Supplemental Figure 3. Endocytosis of R9 via nucleation zones. Medium containing R9 at a concentration of 20 µM was added to HeLa cells and uptake of fluorescence was followed by time lapse confocal microscopy. The time series was started 1 minute after addition of peptide. The video comprises 50 images with a time interval of 10 seconds between each image.

Video 1 (mov)