4.7. F-Actin Staining Using TRITC-Conjugated Phalloidin

YD-15 cells were seeded on coverslips and treated with DMSO, NCTD, or PF562271 for 24 h. The cells were fixed and permeabilized with Cytofix/Cytoperm (BD Bioscience) for 40 min at 4 °C and incubated with TRITC-labelled phalloidin for 40 min, washed with PBS containing 0.1% BSA, and then reacted with 5 mg/mL 4,6-diamidino-2-phenylindole (DAPI; Sigma) for nuclei staining. After washing, the samples were rinsed with PBS and mounted with a mounting media. Immunofluorescence images were obtained using LSM700 confocal laser scanning microscope (Carl Zeiss, Oberkochen, Germany).