Experiment 4: Klf4, FoxD3, FoxD5 and combined FoxD3 and FoxD5 loss-of-function experiments at 30% epiboly Wild Type zebrafish embryos
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ABSTRACT: Morpholino injections were performed to investigate transcriptional targets of Klf4, FoxD3 and FoxD5 in the blastula stage embryos (4,7 hours postfertilization) of wild type zebrafish embryos. Experiments were performed in biological duplicates or triplicates. Cy3 and Cy5 channels were splitted in Genedata Analyst software, quantile normalised and used for independent comparisons.
Project description:Morpholino injections were performed to investigate transcriptional targets of Klf2a, and Klf2b in the blastula stage embryos (4,7 hours postfertilization) of wild type zebrafish embryos. Experiments were performed in biological duplicates. Cy3 and Cy5 channels were splitted in Genedata Analyst software, quantile normalised and used for independent comparisons.
Project description:Overexpression experiments with or without protein synthesis inhibition were performed to investigate transcriptional targets of Klf2a, Klf2b and FoxD3 in the blastula stage embryos (4,7 hours postfertilization) of wild type or MZspg backgroumd Experiments were performed in biological duplicates. Cy3 and Cy5 channels were splitted in Genedata Analyst software, quantile normalised and used for independent comparisons.
Project description:Pou5f1 and Sox2 overexpression experiments with protein synthesis inhibitor were performed to investigate direct transcriptional targets of Pou5f1 and Sox2 Experiments were performed in biological triplicates. Cy3 and Cy5 channels were split and analyzed separately using Genedata Analyst software, quantile normalised and used for independent comparisons.
Project description:Overexpression experiments with or without protein synthesis inhibition were performed to investigate transcriptional targets of Klf4, FoxD3, and Nanog-like in the blastula stage embryos (4.7 hours postfertilization) of wild type or MZspg background. Experiments were performed in biological triplicates. Cy3 and Cy5 channels were splitted in Genedata Expressionist and Analyst software, quantile normalised and used for independent comparisons.
Project description:Analysis of transcriptional profiles in Sbds(ATG) MO-injected embryos with and without coinjection of p53(ATG) MO. We identified a large number of changes in transcript abundance associated with loss of Sbds. Among the 24,278 annotated zebrafish genes in the platform, 4,892 significantly differentially expressed genes were identified. Embryos were carefully staged at 24 hpf and RNA was extracted from approximately 50 embryos per condition in Trizol, followed by purification using RNA miniprep columns (Qiagen). Four independent experiments were extracted. cDNA microarrays were performed at the Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins Microarray Core Facility using 4x44K Zebrafish gene expression microarray slides (Agilent, Santa Clara, CA).
Project description:Total transcriptome analyses were carried out to investigate changes in transcript patterns in the Mycobacterium smegmatis wild type strain SMR5 due to nitrogen starvation and to expand the knowledge about the role of the two transcriptional regulators of nitrogen metabolism, namely GlnR and AmtR, in these processes. A first experiment revealed enhanced transcript levels of 284 genes and reduced transcripts of 231 genes in the wild type under nitrogen starvation compared to nitrogen surplus. When glnR deletion strain MH1 was compared to the wild type under nitrogen starvation, decreased transcript levels of 125 genes were detected, indicating that these are activated by GlnR due to nitrogen limitation. Comparing amtR deletion strain YL1 to the wild type under nitrogen starvation, enhanced transcript levels of 2 genes were found, indicating that they are repressed by AmtR under nitrogen surplus. A comparison of YL1 and wild type under surplus, as well as a comparison of YL1 under nitrogen surplus and starvation and of MH1 under nitrogen surplus and starvation were carried out as additional control experiments. It can be concluded that GlnR is the master regulator of nitrogen control in M. smegmatis and that AmtR fulfills only a small, subordinate role in the regulation of an operon. This SuperSeries is composed of the following subset Series: GSE30033: Mycobacterium smegmatis - Comparison of glnR deletion strain MH1 under nitrogen surplus and starvation GSE30231: Mycobacterium smegmatis - Comparison of wild type SMR5 and amtR deletion strain YL1 under nitrogen starvation GSE30232: Mycobacterium smegmatis - Comparison of amtR deletion strain YL1 under nitrogen surplus and starvation GSE30233: Mycobacterium smegmatis - Comparison of wild type SMR5 under nitrogen surplus and starvation GSE30234: Mycobacterium smegmatis - Comparison of wild type SMR5 and glnR deletion strain MH1 under nitrogen starvation GSE30235: Mycobacterium smegmatis - Comparison of wild type SMR5 and amtR deletion strain YL1 under nitrogen surplus Refer to individual Series
Project description:A zebrafish model of arterial tortuosity syndrome (ATS) was generated by knocking down the slc2a10/glut10 gene using antisense morpholino oligonucleotides (MO). Control morpholino treated embryos were used as controls. The samples were collected for gene expression profiling at 48 hours post fertilization. Experimental details and analyzed data are available in Willaert et al. Human Molecular Genetics 2011; doi: 10.1093/hmg/ddr555 Two-condition experiment, slc2a10 MO (7.5ng) treatment vs control MO (5ng) treatment. Biological replicates: 3 slc2a10 MO replicates, compared to a pooled sample of 3 control MO replicates with dye swap.
Project description:This SuperSeries is composed of the following subset Series: GSE34508: Knockdown of slc2a10/glut10 in zebrafish embryos vs. controls GSE34509: TGFb receptor 1 (TGFBR1) inhibitor treatment of zebrafish embryos vs. controls Refer to individual Series