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RNAseq of isolated mitotic HeLa cells treated with Actinomycin D

ABSTRACT: RNAseq of isolated mitotic HeLa cells treated with Actinomycin D

PROVIDER: PRJEB27865 | ENA |

REPOSITORIES: ENA

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RNAseq of isolated mitotic HeLa cells treated with Actinomycin D

Project description:The goal of this experiment was to assess whether transcription happens during mitosis. For this, we isolated mitotic HeLa cells and treated them with a transcription inhibitor, ActD.

2018-08-28 | E-MTAB-6661 | biostudies-arrayexpress

RNA half-life analysis after transcriptional blocking with actinomycin D

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RNA half-life analysis after transcriptional blocking with actinomycin D

Project description:Transcriptional profiling of HeLa cells treated with actinomycin D for increasing amounts of time. Relative abundance to control cells was used to estimate the half-life of lncRNAs. Two-condition experiment, actinomycin D-treated vs. untreatted HeLa cells. Biological replicates: 2. Technical replicates: 2.

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Expression profiling of HeLa cells at S and G2/M phases of mitotic cell cycle.

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Transcription profiling of cell cycle in HeLa S3, mitotic selection, experiment 1

Project description:HeLa S3 cells in mitosis were selected by mitotic shake-off using an automated cell shaker (Eliassen et al. 2000, MCB)by Beth Alexander and Myra Hurt at Florida State University. This experiment set contains the complete set of 43K arrays for the first mitotic shake-off experiment described in Whitfield et al.(2002), (Shake).

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Mitotic Hela cells expressing GFP-Mitotic Hela cells expressing GFP-Polθ vs Mitotic Hela cells expressing GFP-Polθ treated with Volasertib vs Mitotic Hela cells expressing GFP-Polθ treated with Volasertib

Project description:We show that Polθ is recruited to mitotic Double-strand breaks (DSBs) to slow down cell cycle progression and allow DNA repair. Because Polθ is one of the only repair protein to forms repair foci during mitosis, we investigated its regulation during mitosis. We performed immunoprecipitation (IP) of Polθ and assessed phosphorylation by immunoblot analysis (using pan phospho antibodies). We observed a phosphorylation signal corresponding to the size of Polθ when IP was performed from mitotic cell extracts. This phosphorylation signal was abolished when cells where treated with two different PLK1 inhibitors (PLK1i), indicating that PLK1 is responsible for Polθ phosphorylation in mitosis. In order to elucidate the regulation of mitotic Polθ activity, we performed mass spectrometry (MS)-based phosphorylation analysis of Polθ in mitosis with or without the PLK1 inhibitor Volasertib. We found 5 phosphorylated residues. To assess the functional consequences of Polθ phosphorylation by PLK1, we mutated some of the identified residues and found that the phospho dead mutant of Polθ fails be recruited to DSBs in mitosis. This indicates that PLK1-mediated regulation of mitotic Polθ repair is essential for its proper functioning

2023-09-15 | PXD036486 | Pride

Hela Mitotic Selection - gene expression time series in early G1 phase of the human cell cycle

Project description:HeLa mitotic cells were collected using a mitotic shakeoff apparatus to study gene expression in early G1 phase of the human cell cycle. This set includes microarray data from two shake-off experiments : shake 1 (0h-14h) which was conducted over 14 hours and RNA samples collected every 2 hrs, and shake 2 (0min-120min) which was conducted over 2 hours and samples collected every 15 minutes. Total RNA was prepared using ULTRASPEC RNA isolation system, and Reference RNA was extracted from asynchronously growing HeLa cells using TRIzol. For cDNA synthesis and microarray hybridization, refer to Whitfield ML. et al., Mol Biol Cell, 2002. 13(6): p. 1977-2000. Groups of assays that are related as part of a time series. Age: g1 progression after mitotic shake-off Replicate: two biological replicates (shake1 and shake2) Keywords: time_series_design

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Hela Mitotic Selection - gene expression time series in early G1 phase of the human cell cycle

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Expression profiling of HeLa cells at S and G2/M phases of mitotic cell cycle.

Project description:We compared the mRNAs expression profile of HeLa cells between two phases of the mitotic cell cycle: S and G2/M phases. Results provide insight into the regulation of transcript levels during mitotic cell cycle progression. HeLa cells were synchronized with double thymidine blockade (12 hours with 2 mM thymidine, 12 hours release, and 12 hours with 2 mM thymidine), and cells were taken after 2 hours release (S phase) and 8 hours release (G2/M phase). Keywords: time course Comparison of mRNAs measured at S phase (2 hours release after double thymidine blockade) and at G2/M phase (8 hours release after double thymidine blockade); 3 biological replicates at each of the two time points; two technical replicates with dye swapping per comparison; additional comparison between G2/M (8h) and S (2h).

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