Comparison of Ets2-null and control ES cell derived lineages
Ontology highlight
ABSTRACT: Ets2-null ES cells are defective in differentiating into cardiac myocytes, evidenced by the lack of spontaneous beating, cardiac transcription factors and structural proteins. With the Phalanx mouse Onearray v2, we compared the gene expression profiles of cells derived from Ets2-null and wildtype ES cells. Standard embryoid body culture protocol was used to induce differentiation of murine ES cells. On the 10th day, replicates of either Ets2-null or control wildtype EB culture were collected in Trizol for total RNA extraction. Microarray was preformed by Phalanx.
Project description:Ets2-null ES cells are defective in differentiating into cardiac myocytes, evidenced by the lack of spontaneous beating, cardiac transcription factors and structural proteins. With the Phalanx mouse Onearray v2, we compared the gene expression profiles of cells derived from Ets2-null and wildtype ES cells.
Project description:We explored the role of mammalian ETS1/2 and Mesp homologues of cardiogenic transcription factors of Ciona intestinalis, to convert primary human dermal fibroblasts into cardiac progenitors. ETS1/2 and Mesp homologues of cardiogenic transcription factors of Ciona intestinalis, to convert primary human dermal fibroblasts into cardiac progenitors. Here we show murine Ets2 has an obligatory role for directing cardiac progenitors during cardiopoesis in embryonic stem cells. ETS2 converted fibroblasts into KDR/Flk1+ replicative cells but, like the purported cardiac master regulatory gene Mesp1, could not by itself generate cardiac progenitors de novo from fibroblasts. Co-expression of both Ets2 and Mesp1, however, successfully reprogrammed differentiated fibroblasts into cardiac progenitors, as shown by the de novo appearance of core cardiac transcription factors, gap junction proteins, sarcomeric proteins, electrical activity and contractility. ETS2 and Mesp1 sit at the pinnacle of the cardiopoesis regulatory hierarchy and are well suited for treating human heart disease. Co-expression of both Ets2 and Mesp1, reprogrammed differentiated fibroblasts into cardiac progenitors All sample were done in triplicates, controls were NHDF and ETS2 only infected cells. NHDF were first infected with Doxycyline redulated (Doxy-) ETS2 lentivirus and supplemented with doxycycline for 1 week, sequentially cells were infected with Doxy-Mesp1 and treated for 1 more week. Cells were then aggegated to form EB and hangdrop for 1 week, at the end of that period cells were plated and samples were taken every 24 hrs
Project description:In this study, we identified a number of genes whose expression are regulated by the Hippo tumor suppressor pathway. To disrupt Hippo signaling in the mouse heart, we inactivated the single mammalian Salv ortholog using a Salv conditional null allele and the Nkx2.5 cre allele that directs cardiac cre activity. Total RNA from embryonic hearts were used for expression profiling of 17,455 unique genes. Genes transcriptionally regulated by the Hippo pathway during cardiogenesis were identifed through expression profiling of 17,455 unique genes in Salvador mutant (Salv CKO) embryonic hearts. Total RNA from E9.5-stage hearts were pooled into biological replicate samples (2 control and 2 Salv CKO) and were used to generate expression profiles.
Project description:Current study will help us to elucidate the differential gene expression profiles of two different subsets of monocytes stimulated with gamma-M. tuberculosis CD14hiCD16- and CD14loCD16+ monocytes were isolated from human peripheral blood using magnetic beads for CD14 and CD16. 1.5 million cells were cultured in 1 ml of complete RPMI medium and stimulated with 10 microgram/ml of gamma-M. tuberculosis for 48 hours. After 48 hours cells were harvested and RNA was isolated from control and stimulated samples from CD14hiCD16- and CD14loCD16+ monocytes. So total we had 4 samples: 1. Control CD14hiCD16- 2. Stimulated CD14hiCD16- 3. Control CD14loCD16+ monocytes and 4. CD14loCD16+ monocytes. RNA from all these above 4 samples were sent to Phalanx Biotech Group for whole genome microarray.
Project description:We explored the role of mammalian ETS1/2 and Mesp homologues of cardiogenic transcription factors of Ciona intestinalis, to convert primary human dermal fibroblasts into cardiac progenitors. ETS1/2 and Mesp homologues of cardiogenic transcription factors of Ciona intestinalis, to convert primary human dermal fibroblasts into cardiac progenitors. Here we show murine Ets2 has an obligatory role for directing cardiac progenitors during cardiopoesis in embryonic stem cells. ETS2 converted fibroblasts into KDR/Flk1+ replicative cells but, like the purported cardiac master regulatory gene Mesp1, could not by itself generate cardiac progenitors de novo from fibroblasts. Co-expression of both Ets2 and Mesp1, however, successfully reprogrammed differentiated fibroblasts into cardiac progenitors, as shown by the de novo appearance of core cardiac transcription factors, gap junction proteins, sarcomeric proteins, electrical activity and contractility. ETS2 and Mesp1 sit at the pinnacle of the cardiopoesis regulatory hierarchy and are well suited for treating human heart disease. Co-expression of both Ets2 and Mesp1, reprogrammed differentiated fibroblasts into cardiac progenitors
Project description:In this study, we identified a number of genes whose expression are regulated by the Hippo tumor suppressor pathway. To disrupt Hippo signaling in the mouse heart, we inactivated the single mammalian Salv ortholog using a Salv conditional null allele and the Nkx2.5 cre allele that directs cardiac cre activity. Total RNA from P8-stage hearts were used for expression profiling. Genes transcriptionally regulated by the Hippo pathway during cardiogenesis were identifed through expression profiling in Salvador mutant (Salv CKO) neontate hearts. Total RNA from postnatal day 8 (P8)-stage hearts was purified to generate biological replicate samples (2 control and 2 Salv CKO) and generate expression profiles. 3 technical replicates were used for each sample.
Project description:Microarray analysis of whole lung tissue from three mouse models of asthma Acute asthma was produced by immunization twice, one week apart, of Balb/C mice 8-12 weeks of age with Aspergillus species (5 μg) in adjuvant (1:1 vol/vol). Adjuvant was aluminum and magnesium hydroxide (Pierce). Asthma was initiated by three consecutive intranasal exposures to Aspergillus species (5μg in 15μL saline) and asthma was evaluated 72 hours after the final exposure. Tolerant asthma was produced by intranasal delivery of Aspergillus species (5μg in 15μL saline) twice a week for six consecutive weeks to Balb/C mice 8-12 weeks of age. Asthma was evaluated after mice were rested for an additional three weeks. Chronic asthma was produced by intranasal delivery of the Dustmite/ Ragweed/ Aspergillus (5,50,5ug in 15ul saline) mixture twice a week for six consecutive weeks in female mice Balb/C mice 8-12 weeks of age. Asthma was evaluated 3 weeks after cessation of allergen exposure. Saline control mice were treated intransally with 15ul of saline twice a week for 6 weeks. Asthma was assesed 3 weeks after the final exposure. n=3 samples/ mouse model.
Project description:Conditional Bmp4 overexpression (Bmp4 OE), using a tetracycline regulated Bmp4 gain of function allele, resulted in facial skeletal changes that were most dramatic after an E10.5 Bmp4 induction. We identified a number of genes predominantly transcriptional regulators controlling self-renewal, osteoblast differentiation, and negative Bmp autoregulation whose expression were change after Bmp4 overexpression in the cranial neural crest cells. We performed expression profiling using RNA extracted from E11.5 Bmp4 OE mandibles that were induced with dox for 24 hours. Using a 2-fold change (p< 0.05) as threshold, we identified 144 down-regulated and 120 up-regulated genes.
Project description:Human lactoferrin (LF) is a multifunctional protein involved in immunomodulation, cell growth, and differentiation. In addition to the secreted form (sLF), an alternatively spliced form (ΔLF) that lacks the signal sequence and downregulated in cancer was found. This study was carried out to identify and compare signaling networks provoked by the two LF isoforms. To do this, the two forms were overexpressed in HEK293 cells using the flp-in/tet-on system and genome-wide expression analysis of 18,367 genes was conducted. Secreted form (sLF) or alternatively spliced form (ΔLF) were overexpressed in HEK293 cells using the flp-in/tet-on system and genome-wide expression analysis of 18,367 genes was conducted.
Project description:Transcriptional profiling of ethanol tolerant strains Ets2 and Ets3 comparing control Saccharomyces cerevisiae L3262 with ethanol tolerant strains Ets2 and Ets3, through screening a mutant library of SPT15 of Saccharomyces cerevisiae L3262. Four-condition experiment, L3262 vs. Ets2 or Ets3 strains. Biological replicates: 4 control, each 2 transfected(Ets2, Ets3), independently grown and harvested. One replicate per array.