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HeLa EGFP-Histone H2B Cell Line

Invented by Prof Francis Barr at University of Liverpool
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©2010 Zeng et al. J Cell Biol. Dec 27;191(7):1315-32. doi: 10.1083/jcb.201008106. HeLa cells stably expressing EGFP-tagged histone H2B were transfected with control or protein phosphatase 6 catalytic subunit (PPP6C) si08 duplexes for 48 h and then imaged. An image stack of 25 planes spaced 0.7 μm apart was taken at four stage positions every minute for 10–12 h. Exposure times were 10 ms for EGFP-tagged histone H2B using 1% of full lamp intensity. A brightfield reference image stack was also collected using 10-ms exposures. Arrowhead marks abnormal spindle pole and lagging chromosomes. Bar, 10 μm.
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Catalogue Number 152940
Parental Line HeLa
Synonyms Histone H2B
Host Human
Tissue Cervix
Model Reporter
Relevance The human histone H2B gene was fused to the gene encoding the enhanced green fluorescent protein (EGFP) and transfected into human HeLa cells to generate a stable line constitutively expressing H2B-GFP. The H2B-GFP fusion protein was incorporated into nucleosomes without affecting cell cycle progression. The cell line allows high-resolution imaging of both mitotic chromosomes and interphase chromatin.
Research Area Cell Cycle, Cell Type or Organelle Marker, Cell Structure and Motility, DNA Damage and Repair, Genetic Studies Tools
Recommended Growing Conditions DMEM, 10% FBS, 5% CO2, 37oC. Antibiotic resistance for selection of GFP positive cells: 4μg/mL Blasticidine, expression of GFP is quite stable but selecting at least every two passages is recommended.

References

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References: 2 entries

Zeng et al. 2010. J Cell Biol. 191(7):1315-32. PMID: 21187329.

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Protein phosphatase 6 regulates mitotic spindle formation by controlling the T-loop phosphorylation state of Aurora A bound to its activator TPX2.

Europe PMC ID: 21187329

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References: 2 entries

Zeng et al. 2010. J Cell Biol. 191(7):1315-32. PMID: 21187329.

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Protein phosphatase 6 regulates mitotic spindle formation by controlling the T-loop phosphorylation state of Aurora A bound to its activator TPX2.

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