Cells were mounted on glass slides and fixed in 4% paraformaldehyde. The cells were then permeabilized with phosphate-buffered saline (PBS) containing 0.1% Triton X-100 for 15 min and blocked with 1% bovine serum in PBS for 30 min at room temperature. The cells were then incubated with various primary antibodies overnight at 4°C and detected by Alexa Fluor conjugated secondary antibodies (Alexa 488 and Alexa 594; 1:500, Life Technologies) for 1 hour at room temperature in the dark. The cells were then costained with 4′,6-diamidino-2-phenylindole (DAPI; 1:10,000 dilution) to visualize the nuclei. Images were captured under a confocal laser scanning microscope system (Carl Zeiss AG). A representative image for each condition is shown.

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