Project description:Gene expression profiling comparisons of HepG2.2.15 or PLC/PRF/5 cells either mock (M) transfected or transfected with 0.2 microM S2 RNA or Scrambled (SCR) siRNA were carried out in duplicate 48 hours after transfection. The experiments were carried out in duplicate (a and b). The following combinations of RNA were used on 2 slides each: 1. 2.2.15 cells: mock transfection (reference) versus S2 treatment (test) 2. 2.2.15 cells: mock transfection (reference) versus Scr treatment (test) 3. 2.2.15 cells: Scr treatment (reference) versus S2 treatment (test) 4. PLC/PRF/5 cells: mock transfection (reference) versus S2 treatment (test) 5. PLC/PRF/5 cells: mock transfection (reference) versus Scr treatment (test) 6. PLC/PRF/5 cells: Scr treatment (reference) versus S2 treatment (test) A stimulus or stress experiment design type is where that tests response of an organism(s) to stress/stimulus. e.g. osmotic stress, behavioral treatment Keywords: stimulus_or_stress_design
Project description:Parental flies (y1v1;P{TRiP.JF01138}attP2 and y1w*;P{Act5C-GAL4}25FO1/CyO,y+) were mated to obtain progeny with genotypes for ubiquitous DmSTING RNAi and sibling controls. 2-7 day old flies were used for all infections. 23 nl of listeria monocytogenes (strain 10403S) at 1e7 CFU/ml was injected intrathoracically. Mock inoculation was performed with PBS. Whole flies were homogenized 24 h post-infection.
Project description:To assess the mechanisms by which FACT depletion leads to increased sensitivity of cells to be reprogrammed, we measured the chromatin accessibility landscape using ATAC-seq following mock treatment, SSRP1 knockdown, or SUPT16H knockdown in human fibroblasts and mock, hmg-3 or hmg-4 knockdown in whole worms, and differential gene expression in hmg-3 knockout mutants or following mock, hmg-4, or spt-16 knockdown by RNAseq.
Project description:Parental flies (y1v1;P{TRiP.JF01138}attP2 and y1w*;P{Act5C-GAL4}25FO1/CyO,y+) were mated and 4-7 day old flies were used for all subsequent experiments. dSTING RNAi flies expressed the dsRNA hairpin targeting dSTING and had straight wings, while sibling flies not expressing dSTING dsRNA had curly wings. Pathogens used for inoculation were Listeria monocytogenes, Drosophila C Virus (DCV), and Insect Iridescent virus 6 (IIV6), at the following concentrations: 1e7 CFU/ml, 5e7 TCID50/ml, 5e6 TCID50/ml. Mock inoculation was performed with PBS. Whole flies were homogenized 24 or 72 h post-infection.
Project description:To comprehensively understand the characterization of bluetongue virus (BTV)-host interactome, and BTV infection and pathogenic mechanisms, RNA-seq was performed with BTV serotype 1 Y863 strain-infected and mock-infected sheep embryonic testicular cells (OA3.Ts) at 24 hours post-infection.
Project description:To comprehensively understand the characterization of bluetongue virus (BTV)-host interactome, and BTV infection and pathogenic mechanisms, RNA-seq was performed with BTV serotype 1 Y863 strain-infected and mock-infected sheep embryonic testicular cells (OA3.Ts) at 24 hours post-infection.
Project description:We used an in vitro model to study the impact of hantavirus infection on the cellular gene expression profile. To do so, A549 cells were infected with pathogenic DOBV genotypes Dobrava, Kurkino, and Sochi and less-pathogenic TULV . A549 cells were chosen because of their high susceptibility towards hantavirus infection and because many fundamental studies on hantavirus biology and on host gene expression changes following infection have been performed with this cell line. Cells were infected at a high multiplicity of infection of 5 focus forming units (FFU) per cell to guarantee uniform infection of all cells. Total RNA from infected and mock-infected control cells was isolated at 12 h post infection. This point of time was chosen to allow enough time for establishment of infection and progression to early viral gene expression but to avoid the risk of missing gene expression changes associated with the onset of the cellular innate immune response towards infection. The distinct modulation of cellular transcription of A549 cells was analyzed by means of a whole genome cRNA microarray. All experiments were performed in duplicate using RNA samples from two independently infected cell cultures for each analysis. To compare the gene expression profiles, ratios were calculated by dividing the merged normalized signal intensities of infected samples by mock-control signal intensities. Genes that exhibited a ≥2-fold change (FC) in gene expression and signal intensities that were significantly above the background with p-values ≤ 0.01 were chosen for further analysis. The successful infection of A549 cells with all viruses and the uniform progression of infection were confirmed by immunofluorescence microscopy of cells infected in parallel. At 12 h post infection with each virus nearly all cells were infected without showing cytopathic effects. Since different clinical courses are caused by infection with distinct DOBV genotypes, genotype-specific regulation of cellular transcripts were investigated that may be crucial for pathogenesis in vitro. Selected infection regulated candidates were verified by qPCR at different time points after infection.
