Project description:Murine osteoblast-like cell line MC3T3-E1 subclone 4 (ATCC CRL-2593) (ATTCC, Manassas, USA) were stably transfected using 4D-Nucleofector System (Lonza, Basel, Switzerland) to generate cells that overexpress Hnf4α1, Hnf4α2 or an empty vector (Ctr). ChIPseq analyzes were used to highlight the importance of HNF4a1 and HNF4a2 in osteoblastogenesis.
Project description:Our aim is to explore the effect of Hydroxy-carboxylic Acid Receptor 1 on cardiomyocytes. Neonatal rat cardiomyocytes (NRCMs) were isolated and cultured. Subsequently, NRCMs were treated with the agonist of Hydroxy-carboxylic Acid Receptor 1 (HCAR1), 3Cl-HBA at 40 μM for 48 h(HBA1,HBA2 and HBA3) or DMSO as control(C1,C3 and C3). RNA was extracted using the KAPA RiboErase RNA-Seq kit (Roche, Basel, Switzerland), and was analysed using the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA) for quality control. The libraries were sequenced on an Illumina HiSeq X Ten platform. DESeq2 was used to analyse RNA-seq data. Neonatal rat cardiomyocytes (NRCMs) were isolated and cultured. Subsequently, NRCMs were treated with the agonist of Hydroxy-carboxylic Acid Receptor 1 (HCAR1), 3Cl-HBA (40 μM for 48 h) or DMSO as control. RNA was extracted using the KAPA RiboErase RNA-Seq kit (Roche, Basel, Switzerland), and was analysed using the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA) for quality control. The libraries were sequenced on an Illumina HiSeq X Ten platform.
Project description:Murine osteoblast-like cell line MC3T3-E1 subclone 4 (ATCC CRL-2593) (ATTCC, Manassas, USA) were stably transfected using 4D-Nucleofector System (Lonza, Basel, Switzerland) to generate cells that overexpress Hnf4α1, Hnf4α2 or an empty vector (Ctr). RNAseq and differential expression analyzes were used to highlight the importance of HNF4a1 and HNF4a2 in osteoblastogenesis
Project description:Investigation of whole genome gene expression level changes in the youngest expanded leaves of open-pollinated Golden Delicious seedlings either sprayed with reverse osmosis water or acibenzolar-S-methyl (ASM). Treatment was performed by spraying sprayed to runoff (with a pressurized hand sprayer) with the commercial product Bion 50 WG (Syngenta, Basel, Switzerland; 50% of ASM) prepared in reverse osmosis water at a final concentration of 0.4 g/L. The youngest developped leaf of each seedling was sampled 3 days after the treatment.
Project description:To identify signaling pathways that are differentially regulated in human gliomas, a microarray analysis on 30 brain tumor samples (12 primary glioblastomas (GBM), 3 secondary glioblastomas (GBM-2), 8 astrocytomas (Astro) and 7 oligodendrogliomas (Oligo)) and on 5 glioblastoma cell lines (LN018, LN215, LN229, LN319 and BS149) was performed. Normal brain tissue (NB) and normal human astrocytes (NHA) were used as a control. Kinase expression in each tumor was compared to expression in normal brain and expression values from normal human astrocytes were used as an additional control. Keywords: Kinase expression in each tumor was compared to expression in normal brain and expression values from normal human astrocytes were used as an additional control. Frozen tissue samples of human gliomas and normal brain obtained from the operating room were processed according to the guidelines of the Ethical Committee of the University Hospitals of Basel. Human brain tumor cell lines were derived from human patients. The BS149 cell line was generated at the University of Basel, Switzerland while the “LN” series were a kind gift of Erwin van Meir in Lausanne, Switzerland. Normal human astrocytes (NHAs) were purchased by Cambrex (Walkersville, MD) and cultured according to manufacturer’s recommendations.
Project description:We aimed to investigate the involvement of genes in the down-regulation of KRT19 in Huh7 cell lines. Stably knocked down KRT-19 in Huh7, the corresponding empty plasmid of PLL3.7 treated control cells were applied to extract the total RNA from three samples in each group, and were amplified and transcribed into fluorescent complementary DNA (cDNA). Labeled samples were hybridized to the Human Roche NimbleGen mRNA microarray (Roche, Basel, Switzerland).
Project description:Primary human skeletal muscle cells (hSkMCs) were cultured in growth medium and a fraction of dishes was switched to differentiation medium and to differentiation medium containing 2 x 103 U/ml human recombinant TNF-alpha (Roche Applied Science, Basel, Switzerland), respectively. hSkMCs cells were harvested 24 h (myoblasts day one and myotubes day one without and with 2 x 103 U/ml TNF-alpha, respectively,) after the induction of differentiation. The experiments were performed in triplicates.
Project description:This data displays both known and unknown extra-cellular proteins from 13 species of Lactic Acid bacteria found in the honey-crop of the honeybee Apis. mellifera mellifera. The tryptic peptides from the secreted proteins were run on an Agilent HPLC on a C18 reverse phase column (75 µm x 150 mm, particle size 3 µm). Total run time was 90 min and flow rate 300 nl/min. Buffers used for gradient was 0.1% formic acid in water (buffer A) and 0.1% formic acid in acetonitrile (buffer B). The buffer mixing was 5 min 5% buffer B, followed by 5%-45% buffer B in a linear gradient for 50 min, followed by 45%-80% buffer B in a linear gradient for 5 min. The 80% of buffer B was then kept for 15 min and then rapidly back to 5% buffer B for the final 15 min. The fractions from HPLC were loaded on an LCQ Deca XP Plus Ion trap mass spectrometer (ThermoScientific). Genomic DNA were prepared from all 13 LAB strains depicted earlier and sequenced at MWG Eurofins Operon (Ebensburg, Germany) using Roche GS FLX Titanium technology from Roche (Basel, Switzerland). For each genome a shotgun library was constructed with up to 700,000 reads per segment and was generated by sequencing in 2x half segment of a full FLX+ run. Each genome had an 8 kpb long-paired end library constructed. Approximately 300,000 true paired end reads, sequence tags, and scaffolds with GS FLX+ chemistry using 2x half segment of a full run were generated. Clonal amplification was performed by emPCR in both library types. The sequencing was continued until 15-20 fold coverage was reached. The obtained reads were assembled by the software Newbler 2.6 from Roche (Basel, Switzerland). ORF prediction and automated annotation was performed at Integrated Genomics Assets Inc. (Mount Prospect, Illinois, USA). In ORF prediction three different software were used, GLIMMER, Critica, and Prokpeg. Automated annotation was performed with the ERGOTM algorithms (Integrated Genomics Assets Inc. Mount Prospect, Illinois, USA). The resulting mass spectra-files obtained from the mass spectrometry analysis were searched using MASCOT against a local database containing the predicted proteome of the 13 LAB. We used a cut off Ions score of 38 as a value for determining that the protein was identified. Individual ion scores that were greater than 38 indicated identity or extensive homology (p<0.05) of the protein. Protein sequence similarity searches were performed with software BLASTP in the software package BLAST 2.27+ against a non-redundant protein database at NCBI. Pfam (default database), and InterProScan (default databases). Expressed proteins identified by peptide mass fingerprinting were manually re-annotated.