Volocity demo software

DCs derived from GFP transgenic mice were monitored in a live cell imaging system (PerkinElmer, Massachusetts, USA), and time-lapse micrographs were tracked and analyzed using Volocity Demo software. The generated tracking data were used to plot the cell track and calculate the movement data.

An immunofluo-rescence method was used to observe the protein expression of HERG prior to and following drug treatment. The 293 cells were washed three times with PBS and were subsequently soaked in 4% paraformaldehyde for 30 min at room temperature, followed by washing with PBS a further three times. Following permeabilization with 0.5% Triton X-100 for 20 min, the cells were incubated in antisera against the N- or C-terminal antibody of HERG protein (Alomone, cat. nos. APC-062 and APC-109, respectively; 1:400 dilution), initially at 37°C for 1 h, and then at 4°C overnight. Subsequently, the cells were washed three times in PBS and incubated with antirabbit Cy3-conjugated secondary antibody (Beyotime Institute of Biotechnology, P0183; 1:500 dilution) at 37°C for 30 min, followed by three washes with PBS. Finally, cells were incubated with DAPI (Haoxin Biotech, Inc. or Beyotime Institute of Biotechnology) for 5 min and then were washed three times in PBS. Confocal images were captured using a confocal laser scanning microscope (Zeiss LSM510; Zeiss GmbH, Jena, Germany) and analyzed using Volocity^® Demo software (version 6.0; PerkinElmer, Inc.).