Fluoview ver 2.0a viewer software

At 24 h post-transfection, cells were washed with phosphate-buffered saline (PBS) and fixed in 4 % formaldehyde for 15 min at room temperature. Cells were permeabilized in 0.1 % Triton X-100 in PBS, blocked with 3 % FBS in PBS, and then incubated with primary antibody overnight at 4 °C followed by incubation with secondary antibody conjugated to red fluorochromes. The cell nuclei were stained with 1 μg/ml dihydrochloride (Beyotime, China). Cover slips were mounted upside down on slides with suitable volume of ProLong Gold Antifade Reagent (Invitrogen, USA). The cells were examined using an Olympus IX81 fluorescence microscope equipped with the appropriate filter sets and Olympus FluoView Ver.2.0a Viewer software. The antibodies used were anti-Scribble (Santa Cruz Biotechnology, USA), anti-Goat TRITC labeled IgG antibody (KPL, USA).