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Role of Bmi1 in Shh-mediated shift in gene expression levels

ABSTRACT: We investigated the role of Bmi1 in Shh-mediated shift in gene expression levels

ORGANISM(S): Mus musculus

PROVIDER: GSE19176 | GEO | 2010/01/20

SECONDARY ACCESSION(S): PRJNA120593

REPOSITORIES: GEO

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Transcription profiling of mouse Primary cerebellar granule cell precursors (GCPs) cultures were prepared from P7 Bmi-1-/- or wild type to investigate the role of Bmi1 in Shh-mediated shift in gene expression levels

Project description:We investigated the role of Bmi1 in Shh-mediated shift in gene expression levels Experiment Overall Design: Primary cerebellar granule cell precursors (GCPs) cultures were prepared from P7 Bmi-1-/- or wild type mice as described before {Messer, 1977 #810}. Cells were grown in Dulbecco MEM high Glucose, supplemented with 10% FCS glutamine, 20 mM KCl and penicillinâstreptomycin for 24 hours. Shh (R&D Systems) if appropriate was added for 24 hours to give at final concentration of 3 mg/ml. Experiment Overall Design: We performed 3 biological replicates for both WT and Bmi-1-/- cultures, and 2 biological replicates for both WT and Bmi-1-/- cultures, treated with Shh. Experiment Overall Design: Total RNA was isolated using RNeasy Mini Kit (Qiagen) and reverse-transcribed into double-stranded cDNA with One-Cycle cDNA Synthesis Kit (Affymetrix Inc., P/N 900431, Santa Clara, CA) and labelled biotin. Biotin-labeled cRNA samples were fragmented randomly to 35â200 bp and hybridized to GeneChipÂ® Mouse Genome 430 2.0. An Affymetrix GeneChip Scanner 3000 (Affymetrix Inc., Santa Clara, CA) was used to measure the fluorescent intensity emitted by the labeled target.

2010-02-17 | E-GEOD-19176 | biostudies-arrayexpress

Elucidation of BMI1, RYBP and H2AK119ub interactomes in Glioblastoma

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Polarized Dock Activity Drives Shh-Mediated Axon Guidance

Project description:In the developing spinal cord, Sonic hedgehog (Shh) attracts commissural axons toward the floor plate. How Shh regulates the cytoskeletal remodeling that underlies growth cone turning is unknown. We found that Shh-mediated growth cone turning requires the activity of Docks, which are unconventional GEFs. Knockdown of Dock3 and 4, or their binding partner ELMO1 and 2, abolished commissural axon attraction by Shh in vitro. Dock3/4 and ELMO1/2 were also required for correct commissural axon guidance in vivo. Polarized Dock activity was sufficient to induce axon turning, indicating that Docks are instructive for axon guidance. Mechanistically, we show that Dock and ELMO interact with Boc, the Shh receptor, and that this interaction is reduced upon Shh stimulation. Furthermore, Shh stimulation translocates ELMO to the growth cone periphery and activates Rac1. This identifies Dock/ELMO as an effector complex of non-canonical Shh signaling and demonstrates the instructive role of GEFs in axon guidance.

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Mus musculus

Project description:Role of Bmi1 in Shh-mediated shift in gene expression levels

| PRJNA120593 | ENA

Reprogramming of fibroblasts into induced pluripotent stem cells with Bmi1

Project description:Somatic cells can be reprogrammed into induced pluripotent stem (iPS) cells by Oct4, Sox2, Klf4, plus c-Myc. Recently, Sox2 plus Oct4 were shown to reprogram fibroblasts and Oct4 alone to reprogram mouse and human neural stem cells (NSCs) into iPS cells. Here we report that Bmi1 leads to dedifferentiation of mouse fibroblasts into NSC-like cells and, in combination with Oct4, replaces Sox2, Klf4 and c-Myc during reprogramming fibroblasts to iPS cells. Furthermore, activation of sonic hedgehog signalling (by Shh, purmorphamine, or oxysterol) replaces the effects of Bmi1, and, in combination with Oct4, reprograms mouse embryonic and adult fibroblasts into iPS cells. One-and two-factor iPS cells are similar to mouse embryonic and adult fibroblasts into iPS cells in global gene expression profile, epigenetic status, in vitro and in bibo differentiation into all three ferm layers, as well as teratoma formation and germline transmission in vivo. These data support that fibroblasts can be reprogrammed into iPS cells by Oct4 alone. Total RNAs were isolated from indicated cells and labeled with Cy3. Hybridization was performed once for each sample.

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Reprogramming of fibroblasts into induced pluripotent stem cells with Bmi1

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Bmi1 mutant vs. WT dental epithelia

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