Project description:Analysis of heart of Hypoxia-inducible 2 alpha-deficient mice, specific in cardiomyocytes (Hif2aloxP/loxP Myosin Cre+) subjected to myocardial ishcemia reperfusion. Hif2a belongs to the family of transcription factors, that regulate a transcriptional response to tissue hypoxia. Overall design: Untreated Myosin Cre+ and Hif2aloxP/loxP Myosin Cre+ mice were compared to wMyosin Cre+ and Hif2aloxP/loxP Myosin Cre+ mice with 45 min ischemia followed by 2 hours reperfusion.
Project description:RNA sequencing and subsequent bioinformatics analyses were performed at early reoxygenation stages in HL-1 cardiomyocytes treated or not with BRL37344 Methods: HL-1 cardiomyocytes were subjected to Hypoxia/Reoxygenation (6h/1h) with/without a β3AR agonist (BRL37344 5µmol/L). mRNA profiles were generated by deep sequencing, in triplicate, using Illumina GAIIx.The sequence reads that passed quality filters were quantified using BWA aligned reads using RSEM. qRT–PCR validation was performed using SYBR Green assay. Results: After 6h of hypoxia followed by 1h reoxygenation, 866 genes were differentially expressed upon β3AR stimulation by BRL37344. Among these, 177 were at least 2-fold up or downregulated. Conclusions: Our results show that Hsp70 plays a key role in the cardioprotection afforded by β3AR agonism in cardiomyocytes during the early window of H/R. mRNA profiles from cardiomyocyte subjected to H/R (6h/1h) with/without a β3AR agonist were generated by deep sequencing, in triplicate, using Illumina GAIIx.
Project description:We conducted whole miRNA screening against the MM patients bone marrow (sorted by CD38++) cells that were subjected by chronic hypoxia condition. Overall design: We compired four primary myeloma samples cultured under hypoxia with cultured under normoxia (48hr) to find new important miRNAs of myeloma.
Project description:We conducted whole genome gene screening against the MM patients bone marrow (sorted by CD38++) cells that were subjected by chronic hypoxia condition. Overall design: We compared four primary myeloma samples cultured under hypoxia with cultured under normoxia (48hr) to find new anti-apoptotic gene of myeloma.
Project description:Analysis of murine cardiomyocyte cell line HL-1 treated with Ivermectin or Importazole. Results provide insight into the pathways regulated by the treatments. Overall design: RNA-seq of mouse HL-1 cardiomyocytes treated with vehicle (DMSO), Ivermectin, or Importazole for 24 hours, in triplicate, using Ion Proton System.
Project description:By using world wildly used myeloma cell lines (RPMI8226, KMS-11, KMS-12BM, and MM.1S), we conducted whole miRNA screening against the cells that were subjected by chronic hypoxia condition. Overall design: We compared four myeloma cell lines (RPMI8226, KMS-11, KMS-12BM, and MM.1S) cultured under hypoxia with cultured under normoxia (48hr) to find new important miRNAs of myeloma.
Project description:By using myeloma cell lines (RPMI8226, KMS-11, KMS-12BM, and MM.1S), we conducted whole genome gene screening against the cells that were subjected by chronic hypoxia condition. Overall design: We compared four myeloma cell lines (RPMI8226, KMS-11, KMS-12BM, and MM.1S) cultured under hypoxia with cultured under normoxia (48hr) to find new anti-apoptotic gene of myeloma.
Project description:Introduction: Exosomes are nano-sized extracellular vesicles, released from various cells, which can stimulate or repress responses in target cells. We have recently shown that cultured cardiomyocytes release exosomes and that they, in turn, are involved in facilitating events in target cells by alteration of gene expression. We investigated whether external stimuli of the cardiomyocyte might influence the released exosome characteristics. Material and Methods: Exosomes were isolated from media collected from cultured cardiomyocyte (HL-1) cells with or without growth factor treatment (TGF-beta2 and PDGF-BB), with a series of differential centrifugations. The exosomes were characterized with dynamic light scattering (DLS) and Western blot and analysed with Illumina whole genome microarray gene expression. Results: An average size of 50-80 nm in diameter with no difference between treatment groups was found. Analysis of the mRNA content revealed 623 transcripts in the control group, 691 in the TGF-beta2-treated group and 362 in the PDGF-BB-treated group. 235 transcripts were common for all three groups. Conclusion: We conclude that there is a difference in mRNA content between exosomes derived from cultured cardiomyocytes stimulated with growth factors. We also conclude that all exosomes contain a basic package consisting of ribosomal transcripts and mRNAs coding for proteins with functions within the energy supply system. To study if the transcriptional content in exosomes derived from untreated and growth factor-treated cultured cardiomyocytes (HL-1) differ, and if so, can this difference be explained, 4 control (untreated) exosome samples, 4 TFG-beta2-treated cardiomyocyte-derived exosome samples and 4 PDGF-BB-treated cardiomyocyte-derived exosomes were studied.
Project description:The goal of the study was to identify hypoxia-induced gene expression changes in C. elegans and to determine which of these responses to hypoxia were regulated by hif-1. Towards these aims, we analyzed mRNA expression patterns in synchronized populations of wild-type worms that were cultured in standard lab conditions (normoxia) or in hypoxia. We also assayed mRNA from two mutant strains: (i) animals carrying the strong loss-of-function mutation in hif-1 and (ii) C. elegans that carry a deletion in vhl-1 and express the HIF-1 protein at constitutively high levels. In the microarray experiments, 3 strains were assayed: wild type N2, hif-1 (ia04), and vhl-1 (ok161). Worms were incubated for 4 hours in 21% oxygen or 0.1% oxygen at 210C. Animals were quickly harvested in ice cold M9 buffer, and poly (A) RNA was isolated using established procedures. No more than 3 minutes elapsed between the removal of plates from the hypoxic chamber and the addition of Trizol. RNA was isolated from three independent experiments for each experimental condition (3 genotypes; normoxia vs hypoxia). mRNAs were hybridized to whole-genome microarrays which contained probes for 17,817 predicted genes (94% of the genome). Ch1 in the array represents the data for the worms growed under hypoxia; and Ch2 in the array reprents the data for the worms growed under normoxia. Keywords: Biological Replicates Overall design: Set of arrays that are part of repeated experiments