The lncRNA microarray analysis of 16HBE cell treated with PM2.5 samples in Guangzhou
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ABSTRACT: In order to assess the alteration of lncRNA expression in 16HBE cell treated with PM2.5 samples, we determined the lncRNA expression profiles in 16HBE cell treated with PBS (control group) and PM2.5 samples (low dose 125 μg/mL and high dose 500 μg/mL) using lncRNA Microarray. 16HBE Cells were treated with PM2.5 suspension at concentration of 125 μg/mL and 500 μg/mL, and PBS was used in the control group for 48 h. Then, total RNAs were extracted for lncRNA chip preparation and analysis.
Project description:In order to assess the alteration of lncRNA expression in 16HBE cell treated with PM2.5 samples, we determined the lncRNA expression profiles in 16HBE cell treated with PBS (control group) and PM2.5 samples (low dose 125 μg/mL and high dose 500 μg/mL) using lncRNA Microarray.
Project description:The purpose of this study is to investigate whether the expression profile of long chain non coding RNA (lncRNA) in different concentrations of PM2.5 after treatment of normal human bronchial cells is changed. The total RNAs were extracted from 16HBE cells after exposure to either PM2.5, at concentration of 50 μg/mL and 100μg/mL, or PBS (control group) for 48h. lncRNA-sequencing, detection and analysis was performed. Fold change ≥ 2 and p ≤ 0.05 were used as a cutoff values to determine significant differential expression Further, cluster analyses were performed to classify lncRNAs based on their distinguishable expression in different samples.
Project description:Examine the toxic effect and molecular mechanisms of PM2.5 in primary human umbilical vein endothelial cells (HUVECs) . We used microarrays to detail the global programme of gene expression in HUVECs exposed to PM2.5 and identified distinct classes of up-regulated genes. Cultured HUVECs which were treated with PM2.5 at the concentrations of 50 μg/mL or DMEM for 24 h were collected for RNA extraction and hybridization on Affymetrix microarrays.
Project description:The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. The transcriptional profiles of A. fumigatus conidiospores incubated in the presence or absence of human cells were compared, revealing significant changes in fungal gene expression in response to conidial interaction with cells. Gene expression levels were investigated in A. fumigatus conidiospores incubated in the presence or absence of 16HBE cells for 6 hours. Four independent samples were analysed for each of these conditions, using JCVI PFGRC Aspergillus fumigatus Version 3 microarrays, with a dye-swap experimental design.
Project description:The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. Comparing the transcriptional profiles of control cells and cells exposed to A. fumigatus conidiospores using Agilent Whole Human Genome microarrays revealed significant changes in gene expression in response to the presence of conidiospores. The identification of biologically relevant responses validates this methodology, and motivates further work to characterize the interactions between A. fumigatus conidiospores and primary airway epithelial cells of normal and asthmatic origins. Gene expression levels were investigated in 16HBE cells incubated in the presence or absence of A. fumigatus conidiospores for 6 hours. Four independent cell samples were analysed for each of these conditions, using Agilent Whole Human Genome microarrays, with a one-colour experiemntal design.
Project description:Overnight cultures of S. aureus were diluted to OD600 = 1 and 1 ml of culture was resuspended in 500 μl of either PBS, human plasma, or human serum, and incubated rotating at 37 °C for 30 m. Suspensions were washed 3x with PBS supplemented with 500 40% sucrose and 20 mM Sodium Azide. Cells were incubated with immobilized trypsin (ThermoFisher 20230), suspended in PBS supplemented with 500 40% sucrose and 20 mM Sodium Azide, at 37 °C for 2 h. cells were pelleted, and the supernatant containing protein fragments was analyzed by mass spectrometry to determine proteins.
Project description:The cells of the airway epithelium play critical roles in host defense to inhaled irritants, and in asthma pathogenesis. These cells are constantly exposed to conidiospores of the ubiquitous mould Aspergillus fumigatus, which are small enough to reach the alveoli. This exposure is asymptomatic in most individuals but can be associated with a spectrum of diseases ranging from asthma and allergic bronchopulmonary aspergillosis to aspergilloma and invasive aspergillosis. Airway epithelial cells have been shown to internalize A. fumigatus conidiospores in vitro, but the implications of this process for pathogenesis remain unclear. We have developed a cell culture model for this interaction using the 16HBE cell line and a transgenic A. fumigatus strain expressing green fluorescent protein. Using fluorescence-activated cell sorting (FACS), cells directly interacting with conidiospores and cells not associated with any conidiospores were sorted into separate samples. Transcriptional profiling of these samples using Agilent Whole Human Genome microarrays revealed significant changes in gene expression in response to interaction with conidiospores. The identification of biologically relevant responses validates this methodology, and motivates further work to characterize the interactions between A. fumigatus conidiospores and primary airway epithelial cells of normal and asthmatic origins. Four independent samples of 16HBE cells incubated with GFP-expressing Aspergillus fumigatus conidiospores for 6 hours were analyzed by flow cytometry and divided into negative and positive samples based on the presence of a GFP signal, representing cells with no direct contact with conidiospores and cells directly interacting with conidiospores, respectively. Gene expression was analyzed in these 8 samples using Agilent Whole Human Genome microarrays, with a one-colour experiemntal design.
Project description:Preparation of AMDs. 125 μM Ac4ManNAz were incubated with Raw 264.7 (~ 107 cells/dish) for 24, 48, and 72 h, followed by gentle washing with PBS or complete medium to produce Raw 264.7-Ac4ManNAz. Subsequently, Raw 264.7-Ac4ManNAz (~ 107 cells) were resuspended in PBS containing Alkynyl-Ce6(0.1 μM), ascorbic acid (0.2 μM) and CuSO4 (0.2 μM) and incubated for 1h at 37 °C. Then centrifuged (1000 rpm, 3 min) and washed 3 times with PBS to obtain precursor of AMDs. precursor of AMDs (~ 107 cells/tube, 1.0 ml) was immersed in liquid nitrogen, 24 h later, the frozen tubes were thawed at 37°C, centrifuged at 1500 rpm for 3 min, washed and resuspended by PBS to generate the AMDs (5 x 106 units/tube, 1.0 ml).
Project description:To compare MicroRNA expression in 16HBE infected with EV71 and CA16 we defined the following experimental groups: EV71-0h, EV71-6h, EV71-12h, CA16-0h, CA16-6h and CA16-12h