Project description:Purpose: Despite an overall good outcome of hepatoblastoma patients, prognosis is still poor if associated with metastatic disease. Transcriptomic profiling enables a global view on deregulated cellular pathways. The goal of this study was to compare RNAseq data of metastasized and non-metastasized specimens to identify new molecular players for high-risk hepatoblastoma patients.
Project description:Sp8 and Sp6, members of the Sp family of transcription factors, are required in a dose-dependent manner to induce Fgf8 and En1 in the limb bud ectoderm, therefore controlling proximo-distal and dorso-ventral limb development. Mouse genetics revealed that Sp8 makes a much greater contribution than Sp6 but the nature of its regulatory mechanism was unknown. Here, by combining ChIP-seq and RNA-seq genome-wide analyses we show that Sp8 predominantly functions as an activator from putative distal enhancers regulating crucial limb patterning genes and underscoring its master role in limb development. We also provide compelling evidence for Sp8 cooperating with Dlx5 for the regulation of a considerable set of its target genes. Our work supports a model in which Sp8, Sp6 and Dlx5 act conjointly to regulate target genes with a final functional outcome that depends on their relative availability. This should be considered when interpreting Sp and Dlx mutant phenotypes.
Project description:Sp8 and Sp6, members of the Sp family of transcription factors, are required in a dose-dependent manner to induce Fgf8 and En1 in the limb bud ectoderm, therefore controlling proximo-distal and dorso-ventral limb development. Mouse genetics revealed that Sp8 makes a much greater contribution than Sp6 but the nature of its regulatory mechanism was unknown. Here, by combining ChIP-seq and RNA-seq genome-wide analyses we show that Sp8 predominantly functions as an activator from putative distal enhancers regulating crucial limb patterning genes and underscoring its master role in limb development. We also provide compelling evidence for Sp8 cooperating with Dlx5 for the regulation of a considerable set of its target genes. Our work supports a model in which Sp8, Sp6 and Dlx5 act conjointly to regulate target genes with a final functional outcome that depends on their relative availability. This should be considered when interpreting Sp and Dlx mutant phenotypes.
Project description:Purpose: To asses changes in gene expression profiles from the P11 no cre littermate control olfactory bulbs and conditional double knockout olfactory bulbs of Dlx5/6-CIE; Sp8 Flox/Flox; Sp9 Flox/Flox (Sp8/Sp9-DCKO) mice. Methods: Total RNA was isolated and sequenced from the olfactory bulbs of the P11 no cre littermate controls or Sp8/Sp9-DCKO mice in duplicate using an illumina high-seq 2500. Raw data was analyzed using TopHat. Genes were considered changed which performed fold-change>=2 and FDR<=0.05. Changed genes were then filtered to reveal the downstream targets of Sp8 and Sp9. Results: 32 genes were significantly increased and 144 genes were significantly decreased in expression level due to the loss of Sp9 and Sp8 expression.
Project description:We conducted RNA-Seq and in situ hybridization to identify transcriptional profile changes in the Sp8/Sp9-DCKO olfactory bulb. Simultaneously, we did ChIP-Seq with homemade Sp9 antibody to identify Sp9 binding motifs and which changed genes might be the direct targets of transcription factor Sp9. We found that Sp9 directly binded to key genes which are required for olfactory bulb development. Transcription factors Sp8 and Sp9 belong to the same transcription factor family, so we think Sp8 and Sp9 have very similar binding sites of transcriptional regulation.
Project description:Purpose: To asses changes in gene expression profiles from the P30 wild type littermate control olfactory bulbs and conditional double knockout olfactory bulbs of hGFAP-Cre; Sp8 Flox/Flox; Sp9 Flox/Flox mice. Methods: Total RNA was isolated and sequenced from the olfactory bulbs of the P30 wild type littermate controls or hGFAP-Cre; Sp8 Flox/Flox; Sp9 Flox/Flox mice in tetrad using an illumina high-seq 2500. Raw data was analyzed using TopHat. Genes were considered changed which performed fold-change>=2 and FDR<=0.05. Changed genes were then filtered to reveal the downstream targets of Sp8 and Sp9. Results: 31 genes were significantly increased and 74 genes were significantly decreased in expression level due to the loss of Sp9 and Sp8 expression.
Project description:Purpose: To assess changes in gene expression profiles from the E16.5 no cre littermate control striatum and conditional double knockout of Dlx5/6-CIE; Sp8 Flox/Flox; Sp9 Flox/Flox mice. Methods: Total RNA was isolated and sequenced from the striatum of the E16.5 no cre littermate control or Dlx5/6-CIE; Sp8 Flox/Flox; Sp9 Flox/Flox mice in duplicate using an illumina high-seq 2500. Raw data was analyzed using TopHat. Genes were considered changed which performed fold-change>=2 and FDR<=0.05. Changed genes were then filtered to reveal the downstream targets of Sp8 and Sp9. Results: 80 genes were significantly increased and 120 genes were significantly decreased in expression level due to the loss of Sp8 and Sp9 expression.