Effect of Lycium barbarum polysaccharide-glycoprotein(LBP) on irradiated HaCaT cells
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ABSTRACT: To investigate the protective effect of Lycium barbarum polysaccharide-glycoprotein (LBP) in the radiation-induced HaCaT cell injury, We performed gene expression profiling analysis using data obtained from RNA-seq of HaCaT cells at 3h after radiation.
Project description:This study aims to investigate the binding DNA lesions of AIM2 in HaCaT cells exposed to 0Gy and 20Gy by ChIP-Seq. In this study, HaCaT cells were maintained in DMEM. All culture media were supplemented with 10% fetal bovine serum (FBS; Biological Industries, Kibbutz Beit-Haemek, Israel). Cells were grown at 37 °C in 5% CO2 incubators. Cells were exposed to 20Gy of ionizing radiation using X-ray linear accelerator (Rad Source, Suwanee, GA) at a fixed dose rate of 1.15 Gy/min.Four hours after irradiation, specific antibody against AIM2 was used to pull down AIM2 and the binding DNA lesions, which was then analyzed through ChIP-Seq.
Project description:This study aims to compare mRNA expression between radiated and non-radiated human keratinocyte HaCaT cells by microarray analysis. Human keratinocyte HaCaT cells were divided into two groups, each group has three repeats. The cells were irradiated with a single dose of 0 or 20Gy of X-ray irradiation. 48 hours post radiation, cells from 0 or 20 Gy groups were collected and subjected to microarray analysis. mRNA and lncRNA profilings of each group were analyzed by microarray.
Project description:The p53 protein is encoded by TP53 gene and plays the key role in significant number of cellular processes including proliferation, apoptosis and regulation of many stress response pathways. P53 acts like a direct transcription activator of numerous genes regulating cell cycle arrest, DNA repair, growth inhibition and many others (Mollereau and Ma, 2014). The canonical biological function of p53 is maintaining genome integrity via elimination of damaged or exposed to genotoxic stress cells. Immortalized HaCaT cells are widely used for keratinocyte research, since they maintain stable keratinocyte phenotype, have nearly unlimited proliferative potential, do not require specific growth and differentiation factors (Colombo et al., 2017). Also, HaCaT cells produce typical differentiation markers such as cytokeratins K14 and K10, involucrin (Colombo et al., 2017) and respond to keratinocyte differentiation stimuli. Taking together, HaCaT cells have similar to normal human keratinocytes (NHK) properties, however, as many of spontaneously immortalized cell lines HaCaT cells bear two mutant p53 alleles - R282Q and H179Y (Lehman et al., 1993). Mutp53 in HaCaT has an increased affinity to other p53 family members (p63, p73), which significantly expands p53 properties. Moreover, mutp53 indirectly affects specific target genes via protein-protein interactions with other transcription factors (NF-Y, E2F1, NF-KB) or by tethering p63 to new promotor locations. For more detailed investigation of mutp53 impact on various processes in HaCaT cells we performed a shRNA mediated knockdown of mutp53. For generation of stable TP53 knockdown we employed plasmid vector pLKO-p53-shRNA-941 (Addgene # #25637) followed by puromycin selection of transduced cells. Here we present proteomic dataset obtained from wild type HaCaT cells and p53 knock down HaCaT keratinocytes.
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare transcriptome profiling (RNA-seq) between HaCaT cell and OSM-treated HaCaT cells Methods: mRNA profiles of HaCaT cells and OSM-treated HaCaT cells were generated by deep sequencing, in triplicate, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks.
Project description:Human m6A-mRNA&lncRNA Epitranscriptomic Microarray of arsenite-transformed human keratinocytes (HaCaT-T cells, 1 μM arsenite exposure for 50 passages) compared to its control HaCaT cells (passed for 50 passages without arsenic exposure).
Project description:Analysis of Pseudomonas aeruginosa PA01 (ATCC 15692) Psl polysaccharide deficient mutant. Psl polysaccharide deficient mutant is evaluated with microarray to understand the genes affected by this polysaccharide. Our results provide new vision on the roles played by Psl polysaccharide in P. aeruginosa. Exopolysaccharide Psl is a critical biofilm matrix component in Pseudomonas aeruginosa, which forms a fiber-like matrix to enmesh bacteria communities. Iron has been shown to serve as a signal in P. aeruginosa biofilm development, yet how iron controls biofilm formation is not clear. Here we perform a transcriptomic analysis to compare Psl negative strain versus its isogenic wild-type strain PAO1. The results indicate that the expression of genes involved in iron homeostasis and oxidative stress response increased drastically at transcriptional level in Psl negative strain, suggesting Psl deficiency induces iron limitation. Subsequent studies confirm that Psl can efficiently bind iron in vitro and Psl fibers functions as an iron storage channel in P. aeruginosa biofilms. We used two wild type (WT) replicates and two mutant (MT) replicates to compare the differential gene expression.
Project description:Analysis of Bacteroides thetaiotaomicron (BT) from the ceca of ex-germ free Fut2+ or Fut2- mice on polysaccharide rich or glucose rich polysaccharide deficient diet. BT is involved in the breakdown of plant polysaccharides and is also efficiently utilizes host glycans. BT-colonized mice represent a human gut ecosystem model. Results identify genes that may endow flexibility in adapting to dietary changes by mucosal foraging depending on host fucosylation status In vitro transcriptional profiles of Bacteroides thetaiotaomicron obtained from ceca of ex-germ free Fut2+ or Fut2- mice on polysaccharide rich or glucose rich polysaccharide deficient diet.
Project description:Analysis of Pseudomonas aeruginosa PA01 (ATCC 15692) Psl polysaccharide deficient mutant. Psl polysaccharide deficient mutant is evaluated with microarray to understand the genes affected by this polysaccharide. Our results provide new vision on the roles played by Psl polysaccharide in P. aeruginosa. Exopolysaccharide Psl is a critical biofilm matrix component in Pseudomonas aeruginosa, which forms a fiber-like matrix to enmesh bacteria communities. Iron has been shown to serve as a signal in P. aeruginosa biofilm development, yet how iron controls biofilm formation is not clear. Here we perform a transcriptomic analysis to compare Psl negative strain versus its isogenic wild-type strain PAO1. The results indicate that the expression of genes involved in iron homeostasis and oxidative stress response increased drastically at transcriptional level in Psl negative strain, suggesting Psl deficiency induces iron limitation. Subsequent studies confirm that Psl can efficiently bind iron in vitro and Psl fibers functions as an iron storage channel in P. aeruginosa biofilms.