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Expression of genes in THP-1 macrophages in response to C. glabrata wt, Cgvps34Δ and Cgyps(1-11)Δ cells.

ABSTRACT: To understand the host response of C. glabrata, we have employed the whole genome microarray expression profiling to identify the differentially regulated genes. For this, differentiated THP-1 cells were infected with C. glabrata wt, Cgvps34Δ and Cgyps(1-11)Δ cells for 6 h. Uninfected THP-1 cells, treated with PBS were taken as control. Total RNA was extracted using TriZol Reagent followed by ethonol precipitation. The experiments were performed in biological duplicate.

ORGANISM(S): Homo sapiens

PROVIDER: GSE86176 | GEO | 2018/05/09

REPOSITORIES: GEO

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Project description:A set of 5,908 69-70 mers oligonucleotides that correspond to all C. glabrata putative open reading frames and some non-coding sequences was designed and synthesized at Pasteur Institute, France. Each oilgonucleotide was dissolved into 50% DMSO and spotted on UltraGAPsTM glass slides (Corning Incorporated, Acton, MA) in duplicate to generate whole genome microarray for C. glabrata. The microarray was printed at Biotechnology Research Institute/National Research Council of Canada in Montreal. The sequence information of the oligonucleotides and the micorarray setup can be found at platform GPL3922. Using the microarray, the transcriptional profiles of C. glabrata cells that were coincubated with J774A.1 murine Macrophage-like cells for 2 and 6 hrs were examined. At each time point, three biological repeats were done. 20 ug of total RNA from either macrophage-ingested C. glabrata cells (experimental sample) or the medium-alone controls (control sample) was used to synthesize Cy5- or Cy3-labeled cDNA probes with an anchor primer [(dT)18(dA/dG/dC)] in the presence of Cy5- or Cy3-dCTP (PerkinElmer). For each pair of experimental and control samples, dye-swap was performed, that is, two probe mixtures, the Cy5-experimental plus the Cy3-control probes, and conversely, the Cy3-experimental plus the Cy5-control probes were individually hybridized with a single C. glabrata microarray slide. The hybridization and subsequent washing conditions follows the instruction on the manual for UltraGAPsTM glass slides. The hybridized microarrays were scanned with Axon 4000B scanner, and Data were extracted using Axon GenePix program. Log2 (experimental/control) for each spot on the microarray was calculated. From each dye-swap experiment, the Log2 (experimental / control) for each spot were averaged. Finally, 6 Log2 (experimental/control) values (3 repeats × duplicate spots on each slide) were imported into SAM (significance analysis of microarrays) software for statistical analysis. For genes that are considered significantly induced or repressed, the median of false discovery rate (FDR) equals to 0, while the 90% FDR is less than 0.001. Keywords: transcriptional profiling by microarray

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Expression of genes in THP-1 macrophages in response to C. glabrata wt, Cgvps34Δ and Cgyps(1-11)Δ cells.

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