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Employing a combination of ultraviolet, blue, and green excitation Alexa Fluor (350, 488, and 568, respectively) dyes, a monolayer culture of HJ1.Ov cells was triple-labeled using double immunofluorescence and a phallotoxin. Nuclei were visualized with mouse anti-NPCP (nuclear pore complex protein) primary antibodies, while the Golgi complex was stained with rabbit anti-giantin antibodies. Secondary antibodies were goat anti-mouse and anti-rabbit conjugated to Alexa Fluor 488 and 568. The filamentous actin network was counterstained with Alexa Fluor 350 conjugated to phalloidin.
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