The role of Guanylate-binding protein in Ectromelia virus
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus Musculus
SUBMITTER: 
Zhizhong Jing
PROVIDER: PXD044061 | iProX | Tue Jul 25 00:00:00 BST 2023
REPOSITORIES: iProX
Similar Datasets
Project description:One day before transfection, HeLa cells were seeded in 6-well culture plates (1.5 x 10e5 cells per well) or 10-cm culture dishes (4.3 x 10e5 cells per dish). siRNA duplex (at a final concentration in culture medium of 30 nM) was transfected with Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions. siRNA duplices specific for human hnRNP L, human hnRNP LL, and luciferase GL2 were from MWG Biotech (Ebersberg, Germany). Keywords: control / knockdown comparison
2007-12-01 | GSE8945 | GEO
Project description:The miRNAs were purified from paraffin embedded liver biopsy samples of HCV and HCC patients, using an miRNeasy mini kit (Qiagen) according to manufacturer’s instructions. First, 25mg of liver biopsy tissues was homogenized in 700μl QIAzol lysis reagent, mixed, and shaken with chloroform. After that, the phases were separated by centrifugation at 12000´g for 15 min. The upper aqueous phase was collected in a fresh tube and the remaining DNA and organic protein organic phases was stored at -70°C until required. The RNA aqueous phase was washed further on the column and finally RNA was eluted into DEPC water (Ambion). Purified miRNAs were reverse transcribed into first strand cDNA using a miRNA first strand kit. (Qiagen). The reaction was carried out at 37ºC for 2 hours followed by heating at 95ºC for 5 min.
2019-12-17 | GSE142060 | GEO
Project description:Trypanosomes were sorted (0 cells, 1 cell, 50 cells) using a FACSaria III (BD Biosciences; precision: single-cell; nozzle: 100 µm). Forward-scatter area (FCS-A) versus side-scatter area (SSC-A) was used to gate the cells. Trypanosomes were sorted in 48-wells plate (Brand) filled with 2.6 µL of lysis buffer (0.01 µL of RNAse inhibitor (Takara) and 1x Lysis buffer (Takara) in RNAse-free water). Immediately after sorting cells were placed on ice for 5 minutes and stored at -80 °C. 50 and single trypanosomes were prepared using SMART-Seq v4 Ultra Low Input RNA Kit (Takara) using one fourth of reagents volumes compared to the supplier instructions. PCR amplification was performed using 26 cycles using supplier recommendations. cDNA was purified using XP beads (Beckman Coulter) and recovered in 15 µL of elution buffer (Takara). Libraries were quantified using the Qubit Hs Assay (Life Technologies) and the qualities of the libraries were further monitored using a Bioanalyzer (Agilent). Similar to what has been published previously 19, 1 ng of cDNA was subjected to a tagmentation-based protocol (Nextera XT, Illumina) using one-quarter of the recommended volumes, 10 minuntes for tagmentation at 55 °C and 1 minute extension time during PCR amplification. Libraries were pooled (96 libraries for NextSeq) and sequencing was performed in paired-end mode for 2 × 75 cycles using Illumina's NextSeq 500.
2018-10-09 | GSE120515 | GEO
Project description:Trypanosomes were sorted (1 cell, 10 cells, 50 cells) using a FACSaria III (BD Biosciences; precision: single-cell; nozzle: 100 µm). Forward-scatter area (FCS-A) versus side-scatter area (SSC-A) was used to gate the cells. Trypanosomes were sorted in 48-wells plate (Brand) filled with 2.6 µL of lysis buffer (0.01 µL of RNAse inhibitor (Takara) and 1x Lysis buffer (Takara) in RNAse-free water). Immediately after sorting cells were placed on ice for 5 minutes and stored at -80 °C. 50 and single trypanosomes were prepared using SMART-Seq v4 Ultra Low Input RNA Kit (Takara) using one fourth of reagents volumes compared to the supplier instructions. PCR amplification was performed using 26 cycles using supplier recommendations. cDNA was purified using XP beads (Beckman Coulter) and recovered in 15 µL of elution buffer (Takara). Libraries were quantified using the Qubit Hs Assay (Life Technologies) and the qualities of the libraries were further monitored using a Bioanalyzer (Agilent). Similar to what has been published previously 19, 1 ng of cDNA was subjected to a tagmentation-based protocol (Nextera XT, Illumina) using one-quarter of the recommended volumes, 10 minuntes for tagmentation at 55 °C and 1 minute extension time during PCR amplification. Libraries were pooled (96 libraries for NextSeq) and sequencing was performed in paired-end mode for 2 × 75 cycles using Illumina's NextSeq 500.
2018-10-09 | GSE121026 | GEO
Project description:Frozen PBMC from infants 7 days before randomization (PBMC from two infant were 28 days after randomization) were stimulated in RPMI with 10% Feotal calf serum (FCS) at 5 million/ml with 1 million CFU/ml BCG (SII) or left unstimulated in a total volumn of 200ul/well in 96-well plates. Cells were incubated for 12 hours (37C, 5% CO2). Plates were centrifuged and 200ul supernatant was removed without disrupting the pellet and transferred into clean U-bottom plates. The plates were centrifuged again, the supernatant were flicked-out and the cells were resuspended in 200ul RLT buffer with beta-mercaptoethanol (10ul/ml). The samples were mixed with the buffer to lyse the cells and the plates were sealed and stored at -20C. At the time of RNA-extraction, plates were thawed at 37% for 10 mins. RNA extraction was performed using the Qiagen RNeasy kit following manufacturer's instructions (including optional DNase digestion) and RNA was eluted twice, 30 ul per elution. RNA was quantified by nanodrop and samples were stored at -80C.
2022-05-24 | GSE203395 | GEO
Project description:Purpose:To understand the change in cellular metabolism and function for the overxepression of SCL27A5 in hepatoma cells. Methods:Total RNAs of AdSLC27A5- or AdGFP-infected PLC/PRF/5 cells were extracted using TRIzol (Invitrogen), following the manufacturer’s instructions. RNA-seq and bioinformatic data analysis were performed by Shanghai Novel Bio Ltd. Briefly, strand-specific RNA-seq libraries were prepared using the Total RNA-seq (H/M/R) Library Prep Kit (Vazyme Biotech, Nanjing, China) and were sequenced on a HiSeq X Ten sequencing platform. Raw reads in FASTQ format were subjected to quality control using FastQC. RNA-seq reads were aligned to the reference genome using Bowtie. Uniquely mapped reads were used for further analysis. Gene expression levels are expressed as RPKM (reads per kilobase per million reads) and differences in gene expression were calculated with rSeq. Results:There were 17 genes differentially expressed in AdSLC27A5-infected PLC/PRF/5 cells compare to the GFP control group (fold change >1.5 or < 0.667; FDR < 0.05).
2023-07-01 | GSE117823 | GEO
Project description:0.3g of flowers were ground in liquid nitrogen. The powder was extracted for 30 min [H2] in 1.5 ml of lysis buffer (50mM Tris HCl pH 8.0, 50mM NaCl, 1% Triton x-100 and 1 x cOmplete™ EDTA-free protease inhibitor (Roche)). After removal of cell debris by centrifugation (2 times 10 min, 16000 x g, 4 °C) the cleared supernatants were incubated for 30 min with GFP or FLAG antibodies coupled to magnetic microbeads (μMACS GFP and DYKDDDDK isolation Kits, Miltenyi). Beads were loaded on magnetized MACS separation columns equilibrated with lysis buffer and washed 4 times with 300 µl washing buffer (50mM Tris HCL pH 7.5, 25 or 50 mM NaCl, 0,1 % Triton). Samples were eluted in 50 µl of pre-warmed elution buffer (Milteny). Control IPS were performed in Col-O using either GFP or FLAG antibodies.
2020-04-15 | PXD014447 | Pride
Project description:Expression data from Total RNA extracted from murine spleen. Sepsis was induced in C57Bl/6J mice by cecal ligation and puncture (CLP), followed 6 hours later by an intravenous injection of Mesenchymal Stem Cell (MSC) or saline. Twenty-eight hours after CLP, plasma, bronchoalveolar lavage (BAL) fluid and tissues were collected for analyses. Total RNA was extracted using Trizol (as per manufactures' instruction) followed by clean-up procedure using Qiagen RNA easy Prep (as per manufactures instructions) In the following study we hypothesized that mesenchymal stem cells (MSCs), which have documented immunomodulatory properties, would reduce sepsis-associated inflammation and organ injury in a clinically relevant model of sepsis.
2010-11-22 | GSE24357 | GEO
Project description:Purpose:To understand the change in cellular metabolism and function for the overxepression of GSTZ1 or PCK1 in hepatoma cells. Methods:Total RNAs of AdGFP- , AdGSTZ1-, or AdPCK1-infected Huh7 cells were extracted using TRIzol (Invitrogen), following the manufacturer’s instructions. RNA-seq and bioinformatic data analysis were performed by Shanghai Novel Bio Ltd. Briefly, strand-specific RNA-seq libraries were prepared using the Total RNA-seq (H/M/R) Library Prep Kit (Vazyme Biotech, Nanjing, China) and were sequenced on Ion Torrent Proton sequencing platform. Raw reads in FASTQ format were subjected to quality control using FastQC. RNA-seq reads were aligned to the reference genome using Bowtie. Uniquely mapped reads were used for further analysis. Gene expression levels are expressed as RPKM (reads per kilobase per million reads) and differences in gene expression were calculated with rSeq. Results:There were 498 genes differentially expressed in AdPCK1-infected Huh 7 cells, 513 genes differentially expressed in AdGSTZ1-infected Huh 7 cells compare to the GFP control group respectively (fold change >1.5 or < 0.667; FDR < 0.05).
2019-01-02 | GSE117822 | GEO
Project description:SVF cells were isolated from liposuction material (hips, outer thigh) of healthy controls and lipedema patients and seeded for the collection of cell supernatants after 24 hours. After several centrifugation steps, the supernatants were concentrated and enriched for small extracellular vesicles. Total RNA was extracted from the concentrated supernatants and the small extracellular vesicles fraction. 187 miRNAs were analyzed using qPCR reaction (miRCURY SYBR® Green mastermix, Quiagen).
2020-06-22 | GSE138579 | GEO