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Cdr2 siRNA knockdown during passage through mitosis: HeLa cells, Rat1 wild type and c-myc null cells

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

PROVIDER: PRJNA124295 | ENA |

REPOSITORIES: ENA

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

2010-04-07 | GSE20037 | GEO

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

Project description:[Hela cells]: We performed cdr2 knockdown with a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis. [Rat1a wild type and myc null cells]: We performed cdr2 knockdown using a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis. [HeLa cells]: Cells were transfected with control or cdr2 siRNAs and then cells were synchronized in mitosis using a sequential thymidine/nocodazole block. Cells were subsequently released from nocodazole block and after 3 hours RNA was harvested for microarray analysis [Rat-1 wild type (TGR) and c-myc null (15.19) cells]: Cells were transfected with control or cdr2 siRNAs and then cells were synchronized in mitosis using a sequential thymidine/nocodazole block. Cells were then subsequently released from nocodazole block and after 3 hours RNA was harvested for microarray analysis

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

Attenuation of Zika Virus by Passage in Human HeLa Cells.

Project description:Zika virus (ZIKV) is a mosquito-borne Flavivirus. Previous studies have shown that mosquito-transmitted flaviviruses, including yellow fever, Japanese encephalitis, and West Nile viruses, could be attenuated by serial passaging in human HeLa cells. Therefore, it was hypothesized that wild-type ZIKV would also be attenuated after HeLa cell passaging. A human isolate from the recent ZIKV epidemic was subjected to serial HeLa cell passaging, resulting in attenuated in vitro replication in both Vero and A549 cells. Additionally, infection of AG129 mice with 10 plaque forming units (pfu) of wild-type ZIKV led to viremia and mortality at 12 days, whereas infection with 103 pfu of HeLa-passage 6 (P6) ZIKV led to lower viremia, significant delay in mortality (median survival: 23 days), and increased cytokine and chemokine responses. Genomic sequencing of HeLa-passaged virus identified two amino acid substitutions as early as HeLa-P3: pre-membrane E87K and nonstructural protein 1 R103K. Furthermore, both substitutions were present in virus harvested from HeLa-P6-infected animal tissue. Together, these data show that, similarly to other mosquito-borne flaviviruses, ZIKV is attenuated following passaging in HeLa cells. This strategy can be used to improve understanding of substitutions that contribute to attenuation of ZIKV and be applied to vaccine development across multiple platforms.

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LFQP of lysates from wild-type and TBC1D20-null HeLa cells

Project description:These data are from two wild-type HeLa clones (WT1, WT2) and two TBC1D20-null clones (5H2, 7H5), each analysed in triplicate.

2024-01-26 | PXD016233 | Pride

O-Linked N-Acetylglucosamine Transiently Elevates in HeLa Cells during Mitosis.

Project description:O-linked N-acetylglucosamine (O-GlcNAc) is a dynamic post-translational modification of serine and threonine residues on nuclear and cytoplasmic proteins. O-GlcNAc modification influences many cellular mechanisms, including carbohydrate metabolism, signal transduction and protein degradation. Multiple studies also showed that cell cycle might be modulated by O-GlcNAc. Although the role of O-GlcNAc in the regulation of some cell cycle processes such as mitotic spindle organization or histone phosphorylation is well established, the general behaviour of O-GlcNAc regulation during cell cycle is still controversial. In this study, we analysed the dynamic changes of overall O-GlcNAc levels in HeLa cells using double thymidine block. O-GlcNAc levels in G₁, S, G₂ and M phase were measured. We observed that O-GlcNAc levels are significantly increased during mitosis in comparison to the other cell cycle phases. However, this change could only be detected when mitotic cells were enriched by harvesting round shaped cells from the G₂/M fraction of the synchronized cells. Our data verify that O-GlcNAc is elevated during mitosis, but also emphasize that O-GlcNAc levels can significantly change in a short period of time. Thus, selection and collection of cells at specific cell-cycle checkpoints is a challenging, but necessary requirement for O-GlcNAc studies.

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Early and late passage endothelial cells with or without PDCD10 downregulation

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Expression data of HeLa cells treated with CENP-E siRNA or Eg5 siRNA in the presence of BubR1 siRNA.

Project description:The molecular mechanism responsible for cell fate after mitotic slippage is unclear. We investigated the postmitotic effects of different mitotic aberrations (Ohashi et al. [1]), misaligned chromosomes produced by CENP-E siRNA (siCENP-E), and monopolar spindles resulting from Eg5 siRNA (siEg5) (Miki et al. [2]). To determine which signaling pathways contribute to the postmitotic effect of siCENP-E in the presence of siBubR1 (siCENP-E + siBubR1) compared with siEg5 + siBubR1, we performed comprehensive gene expression analysis using microarray comparisons [1]. The microarray data have been deposited in NCBI's Gene Expression Omnibus (GEO) and are accessible through GEO Series accession number GSE67905.

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The cyclin B1 gene is actively transcribed during mitosis in HeLa cells.

Project description:In mammalian cells, the expression level of the cyclin B1 gene plays a critical role in the progression through mitosis. Here we demonstrate that the transcriptional activity of the human cyclin B1 promoter, as well as the rate of gene transcription, is high during mitosis. Indeed, the cyclin B1 promoter maintains an open chromatin configuration at the mitotic stage. Consistent with this, we show that the cyclin B1 promoter is occupied and bound to NF-Y during mitosis in vivo. Our results provide the first example of RNA polymerase II-dependent transcription during mitosis in mammalian cells.

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Project description:scRNA-seq of E14 Bax null, Atoh7 null, and Bax null/Atoh7 null retinas

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