Fig. 4

PDEP-mediated delivery of an anionic peptide cargo. a Cells were transfected with a plasmid encoding for intact mCherry and non-fluorescent GFP1–10. Cells were incubated, either at room temperature or at 37 °C, with varying concentrations of Peptide-GFP11 for 30 min and analyzed by flow cytometry. Live, single, mCherry-positive cells were first gated. Within this population, mCherry and GFP intensities were measured for about 10,000 cells for each sample. The percentage of GFP-positive cells shown was determined by gating just above background GFP levels. b, c Average GFP intensity for cells treated at 21 °C (b) and at 37 °C (c) with CPP-GFP11 constructs. Averages are divided by the average intensity of cells treated with the GFP11 peptide alone. D-form versions of tat and P14 conjugated to an L-form GFP11 peptide were also analyzed. Error bars are standard deviations of the normalized data, which are shown as open circles, with three data points, from independent experiments, per average