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Cell division cycle in a human cancer cell line (HeLa)

ABSTRACT: This experiment set contains the complete set of raw data for Whitfield et al. (2002)Mol.Biol.Cell. HeLa S3 cells were synchronized by three different methods (double thymidine block, thymidine-nocodazole block or mitotic shake-off)in five independent time courses. mRNA from the first and second double thymidine block experiments were converted to cDNA by standard methods using an oligo-dT/random hexamer mix and hybridized to 24K arrays; mRNA from the third double thymidine and thymidine-nocodazole block experiments were converted to cDNA using olido-dT alone and hybridized to 43K arrays; total RNA from the mitotic shake-off experiment was first amplified using a modified eberwine protocol, aRNA converted to cDNA by reverse transcription with random hexamer and hybridized to 43K arrays. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: Logical Set

ORGANISM(S): Homo sapiens

PROVIDER: GSE3497 | GEO | 2005/10/22

SECONDARY ACCESSION(S): PRJNA93325

REPOSITORIES: GEO

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Cell division cycle in a human cancer cell line (HeLa)

Project description:This experiment set contains the complete set of raw data for Whitfield et al. (2002)Mol.Biol.Cell. HeLa S3 cells were synchronized by three different methods (double thymidine block, thymidine-nocodazole block or mitotic shake-off)in five independent time courses. mRNA from the first and second double thymidine block experiments were converted to cDNA by standard methods using an oligo-dT/random hexamer mix and hybridized to 24K arrays; mRNA from the third double thymidine and thymidine-nocodazole block experiments were converted to cDNA using olido-dT alone and hybridized to 43K arrays; total RNA from the mitotic shake-off experiment was first amplified using a modified eberwine protocol, aRNA converted to cDNA by reverse transcription with random hexamer and hybridized to 43K arrays. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Computed

2005-10-22 | E-GEOD-3497 | biostudies-arrayexpress

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cdr2 siRNA knockdown during passage through mitosis: HeLa cells, Rat1 wild type and c-myc null cells

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2010-04-07 | E-GEOD-20037 | biostudies-arrayexpress

Transcription profiling of cell cycle in HeLa S3 cells, double thymidine block, rxperiment 2

Project description:HeLa S3 cells were synchronized in early S phase with a double thymidine block as described in Whitfield et al(2002)Mol.Biol.Cell. This experiment set contains the complete set of 24K arrays for the second double thymidine block synchronization (Thy-Thy2).

2005-09-22 | E-SMDB-902 | biostudies-arrayexpress

Transcription profiling of cell cycle in HeLa S3 cells, double thymidine block, experiment 3

Project description:HeLa S3 cells were synchronized in early S phase with a double thymidine block as described in Whitfield et al(2002)Mol.Biol.Cell. This experiment set contains the complete set of 43K arrays for the third double thymidine block synchronization (Thy-Thy3).

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Transcription profiling of cell cycle in HeLa S3 cells, double thymidine Block, experiment 1

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Identification of the quantitative phosphoproteome in mitotic HeLa cells with or without G6PD inhibitors.

Project description:HeLa cells were synchronized with a double thymidine block procedure. Briefly, the exponentially growing HeLa cells were maintained with 2 mM thymidine for 18 h, followed by a release of 9 h in fresh medium, and then cells were re-cultured in 2 mM thymidine for additional 15 h. After a release of 7.5 h in fresh medium, DMSO or G6PD inhibitor was added to the medium. 1 h later, the cells entered into M phase. Mitotic cells were harvested by mitotic shake-off. Then, the samples were subjected to LC-MS/MS analysis. Finally, a Kinase-Substrate Enrichment was performed, which could infer the changes of upstream kinase activity upon the treatment of G6PD inhibitors.

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Project description:Flag-G6PD was expressed in HeLa cells. Then the HeLa cells were synchronized with a double thymidine block procedure. Briefly, the exponentially growing HeLa cells were maintained with 2 mM thymidine for 18 h, followed by a release of 9 h in fresh medium, and then cells were re-cultured in 2 mM thymidine for additional 15 h. These interphase cells were harvested. After a release of 8.5 h in fresh medium, the cells entered into M phase. Mitotic cells were harvested by mitotic shake-off. Then, cell lysate was incubated with anti-Flag M2-agarose. Elutes were separated with 10% SDS-PAGE. The gel was stained with Coomassie Brilliant Blue. Visualized bands were excised and subjected to LC-MS/MS analysis.

2023-05-18 | PXD033829 | Pride

Hi-C during mitotic release and G1 entry

Project description:DpnII Hi-C libraries for samples synchronized to G2/M using single 24h thymidine, 3h release, and 12h nocodazole--these samples are then released for x amount of time

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Osteoclast response to low extracellular sodium

Project description:RAW 264.7 murine osteoclastic cells were cultured with recombinant cytokines (RANKL and M-CSF) in culture medium with normal sodium (NS; Na=140 mmol/l) and low sodium and reduced sodium (Na=120 mmol.l) while maintaining normal ostmolality with the addition of mannitol. After 24 hours, total RNA was extracted using PureLink kit (Invitrogen, Carlsbad, CA) according to manufacturer’s instructions, and purified using the Rneasy Mini Kit (Invitrogen). 5 µg RNA from each sample was converted into biotin-labeled cDNA and hybridized to GeneChip Mouse Genome 430 2.0 arrays containing 45,101 probe sets corresponding to known genes and expressed sequence tags. Total RNA was extracted using PureLink kit (Invitrogen, Carlsbad, CA) according to manufacturer’s instructions, and purified using the Rneasy Mini Kit (Invitrogen). 5 µg RNA from each sample was converted into biotin-labeled cDNA and hybridized to GeneChip Mouse Genome 430 2.0 arrays containing 45,101 probe sets corresponding to known genes and expressed sequence tags.

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