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.

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.

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.

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