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Cells were fixed in 2,5% paraformaldehyde (PFA)/PBS for 5 min at room temperature, quenched in 125 mM glycine for 5 min, and subsequently processed for indirect immunofluorescence using standard protocols. Primary antibodies used are described later, and fluorophore-conjugated secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA). DNA was counterstained with 1 μg/ml Hoechst 333342 (Sigma-Aldrich, St. Louis, MO) in PBS and samples mounted onto glass slides with ProLong Gold (Life Technologies).
Copyright and License information: ©2016 Martins This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License ().
“ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
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