Transcriptomics

Dataset Information

5

Transcription profiling of zebrafish embryos injected with a morpholino targeting LIN9


ABSTRACT: TÜAB zebrafish were maintained at 28.5 °C in in 1x Danieau solution (58 mM NaCl, 0.7 mM KCl, 0.4 mM MgSO4, 0.6 mM Ca (NO3)2, 5 mM HEPES, pH 7.6, 0.0001 % Methylene blue). Morpholinos (Gene Tools, Philomath, OR) were designed against Danio rerio lin-9 homolog (Genebank accession NM_001044946). The morpholino (MO) sequences were MO-E1 5´-GTTAGTTTTATTACTCACTCTCGTC-3´ and 5-base mismatch morpholino MO-E1mis 5´-GTTACTTTTAATACTGACTGTCCTC-3´. 7 ng of morpholinos were injected into one cell-stage embryos. 20 embryos per condition (MO E1; MO E1mis) were pooled for RNA purification 24 h post fertilization. Using the two color Quick-Amp Labeling Kit (Agilent; 5190-0444) 100 ng of total RNA were used for cDNA synthesis, mRNA amplification and labeling according to manufacturers instructions. Transcriptional profiling was done on a zebrafish oligo array (Agilent; G2519F AMADID 019161) in a 4 x 44K slide format and analyzed as described before.

ORGANISM(S): Danio rerio  

SUBMITTER: Markus Kleinschmidt  

PROVIDER: E-MEXP-2100 | ArrayExpress | 2009-03-28

REPOSITORIES: ArrayExpress

Dataset's files

Source:
Action DRS
E-MEXP-2100.README.txt Txt
E-MEXP-2100.eSet.r Other
E-MEXP-2100.idf.txt Idf
E-MEXP-2100.idf.txt_original Idf
E-MEXP-2100.processed.1.zip Processed
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Publications

lin9 is required for mitosis and cell survival during early zebrafish development.

Kleinschmidt Markus A MA   Wagner Toni U TU   Liedtke Daniel D   Spahr Susi S   Samans Birgit B   Gaubatz Stefan S  

The Journal of biological chemistry 20090311 19


LIN9 has been described as a regulator of G(1)/S and G(2)/M progression of the cell cycle in invertebrates and human cell lines. To elucidate the in vivo function of LIN9 during vertebrate development, we took advantage of the teleost zebrafish (Danio rerio). By means of antisense morpholinos we show here that Lin9-depleted embryonic cells accumulate in mitosis. Flow cytometry and confocal microscopy data demonstrate that the delay in mitotic progression is followed by apoptosis, which strongly  ...[more]

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