Project description:The erythroid Krüppel-like factor EKLF/KLF1 is a hematopoietic transcription factor binding to CACCC DNA motif and participating in the regulation of erythroid differentiation. With combined use of microarray-based gene expression profiling and promoter-based ChIP-chip assay of E14.5 fetal liver cells from wild type (WT) and EKLF-knockout (Eklf-/-) mouse embryos, we have identified the pathways and direct target genes activated or repressed by EKLF. This genome-wide study together with molecular/ cellular analysis of mouse erythroleukemic cells (MEL) indicate that among the downstream direct target genes of EKLF is Tal1/Scl. Tal1/Scl encodes another DNA-binding hematopoietic transcription factor TAL1/SCL known to be an Eklf activator and essential for definitive erythroid differentiation. Further identification of the authentic Tall gene promoter in combination with in vivo genomic footprinting approach and DNA reporter assay demonstrate that EKLF activates Tall gene through binding to a specific CACCC motif located in its promoter. These data establish the existence of a previously unknow positive regulatory feedback loop between two DNA-binding hematopoietic transcription factors that sustains the mammalian erythropoiesis.
Project description:Homozygous disruption of c-Maf led to embryonic lethality and impaired erythroblastic island formation. c-Maf is expressed in the fetal liver macrophages. It suggests that macrophages are responsible for the lethality of c-Maf knock-out embryos. To search downstream genes of c-Maf, we surveyed genes associated with macrophage function by microarray analysis. keywords: c-Maf, macrophage, erythroblastic islands, WT (c-Maf WT) and c-Maf KO (c-Maf KO) fetal liver macrophages were sorted by a FACSAria cell sorter. Total RNAs from those macrophages were prepared using RNeasy Kit. Genes down-regulated in c-Maf KO macrophages were searched by GeneSpring software.
Project description:The aim of this experiment was to investigate the role of KLF3 in regulating gene expression at different stages throughout the erythroid maturation process. Affymetrix microarrays were performed on fetal liver cells (both TER119- progenitor cells and TER119+ erythroblast cells) from E14.5 wildtype and Klf3 KO mice. Four wildtype TER119- replicates, four Klf3 KO TER119- replicates, four wildtype TER119+ replicates, three Klf3 KO TER119+ replicates. All are from E14.5 fetal liver.
Project description:Investigation of whole genome gene expression level changes in OASIS KO calvaria compared to wild-type calvaria. To gain further insight into the potential mechanisms underlying the defective bone formation in OASIS KO mice, we compared the gene expression in calvaria between WT and OASIS KO mice using a microarray.
Project description:The aim of this experiment was to investigate the role of KLF3 in regulating gene expression at different stages throughout the erythroid maturation process. Affymetrix microarrays were performed on fetal liver cells (both TER119- progenitor cells and TER119+ erythroblast cells) from E14.5 wildtype and Klf3 KO mice.
Project description:Investigation of whole genome gene expression level changes in OASIS KO calvaria compared to wild-type calvaria. To gain further insight into the potential mechanisms underlying the defective bone formation in OASIS KO mice, we compared the gene expression in calvaria between WT and OASIS KO mice using a microarray. Each sample of total RNA was collected from a number of mice.
Project description:The developing erythroid cells require highly coordinated gene expression and metabolism. By comparing the proteomic and transcriptomic changes in human hematopoietic stem/progenitor cells (HSPCs) and lineage-committed erythroid progenitors (ProEs), and uncover pathways related to mitochondrial biogenesis enhanced through post-transcriptional regulation. Two principal mitochondrial factors TFAM and PHB2 are tightly regulated at the protein level and indispensable for mitochondria and erythropoiesis. To determine the role of TFAM in mitochondrial function during erythroid development, we generated Tfam conditional knockout (KO) mice by an erythroid-specific EpoR-Cre allele. We isolated the CD71+Ter119+ embryonic day (E)13.5 fetal liver erythroid cells by FACS sorting, and performed RNA-seq transcriptional profiling analysis.
Project description:Transcriptional profiling of S. thermophilus LMD-9 wild-type compared to the isogenic mutant strain CB052 (deletion of mecA, STER_0216) for the identification of the MecA regulon. Cells were grown in THB medium supplemented with glucose 1% (THBG) and sampled at OD600 = 0.4 for mRNA extraction wild-type vs knockout, 2 WT biological replicates (one used as technical replicate for a dye swap), 2 KO biological replicates (one used as technical replicate for a dye swap)
Project description:Transcriptional profiling of S. thermophilus LMD-9 wild-type compared to the isogenic mutant strain CB003 (deletion of comX, STER_0189) for the identification of the ComX regulon. Cells were grown in CDM medium supplemented with lactose 1% (CDML) and sampled at OD600 = 0.4 for mRNA extraction wild-type vs knockout, 2 WT biological replicates (one used as technical replicate for a dye swap), 2 KO biological replicates (one used as technical replicate for a dye swap)