Figure 2.

Induction of protein modifications with biotin‐cadaverine by nsPEFs. HeLa S3 cells were preincubated with 0.2 mm biotin‐cadaverine for 20 min and subsequently subjected to exposure to nsPEFs. (A) Indicated numbers of shots of nsPEFs at 20 kV·cm⁻¹ were applied to HeLa S3 cells in the presence of biotin‐cadaverine. After 1 h incubation at 37 °C, the cells were subjected to western blot analysis for biotinylated proteins. Ku70 protein is shown as a loading control. Three independent experiments were performed to confirm the reproducibility, and a representative result is shown. (B) HeLa S3 cells were exposed to 35 shots of 20 kV·cm⁻¹ nsPEFs and incubated for the indicated periods. Untreated cells (indicated as –) were included in the analysis. Biotinylated proteins and Ku70 were analyzed by western blotting as described in (A). Three independent experiments were performed to confirm the reproducibility, and a representative result is shown. (C) HeLa S3 cells exposed to nsPEFs (35 shots of 20 kV·cm⁻¹) and untreated control cells were incubated at 37 °C for 1 h and subsequently costained with FITC‐streptavidin, anti‐α‐tubulin antibody, and DAPI. Three independent experiments were performed to confirm the reproducibility, and a representative result is shown. Bar, 20 μm.