Protocol to identify centrosome-associated transcription factors during mitosis in mammalian cell lines

Abstract

Summary: During eukaryotic cell mitosis, the nuclear envelope disintegrates and transcription factors are dissociated from condensed chromosomes. Here, we describe a protocol to study centrosomal translocation of nuclear receptor RXRα. We detail procedures for HeLa cell synchronization followed by immunofluorescence, in situ proximity ligation assay, and centrosome isolation. This protocol can be used to identify other transcription factors associated with the centrosome or other subcellular structures during mitotic progression.For complete details on the use and execution of this protocol, please refer to Xie et al. (2020)

Keywords

• Cell Biology
• Microscopy
• Molecular Biology