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Using double immunofluorescence, an adherent culture of APM cells was washed, fixed, permeabilized, blocked with 10-percent normal goat serum, and treated with a cocktail of rabbit anti-giantin (Golgi complex) and mouse anti-NPCP (nuclear pore complex proteins) primary antibodies. The target antigens were visualized with goat anti-rabbit and anti-mouse secondary antibodies (IgG) conjugated to Alexa Fluor 568 and Alexa Fluor 488, respectively. The filamentous actin cytoskeletal architecture was labeled with Alexa Fluor 350 conjugated to phalloidin.
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