Project description:Gene Set Enrichment Analysis of PEDF knockout livers revealed induction of pathways associated with experimental and human HCC and a transcriptional profile characterized by Wnt/β-catenin activation Genome-wide expression analysis of liver tissue of wild type vs PEDF knockout animals fed high fat diet Total RNA was extracted from whole liver tissue
Project description:Recent studies have reported both mRNA and microRNA (miRNA) in saliva, but little information has been documented on the quality and yield of RNA collected. Therefore, the aim of the present study was to develop an improved RNA isolation method from saliva and to identify major miRNA species in human whole saliva. RNA samples were isolated from normal human saliva using a combined protocol based on the Oragene RNA collection kit and the mirVana miRNA isolation kit in tandem. RNA samples were analyzed for quality and subjected to miRNA array analysis. Twelve representing 6 males and 6 females, were selected for miRNA profiling using the TaqManM-BM-. Low Density Array Card (TLDA) Human miRNA Panel v2.0 (Applied Biosystems). The analysis of expression of the >700 miRNAs was performed by the DNA Core at the Interdisciplinary Center for Biotechnology Research Center at the University of Florida, according to the manufacturerM-bM-^@M-^Ys protocol except the pre-amplification step was omitted. The NormFinder algorithm was used to identify the optimal normalization of miRNA among the 25 most abundantly expressed miRNAs detected.
Project description:We wished to assess the effect of miR196a on overall gene expression in reducing it in high expressing epithelial cells (D19 and B16) using antimiR196a and over expressing it in low-expressing cells (OKF4) using pre-miR196a. We transfected anti miR196a in to D19 and B16 oral keratinocytes which express high levels of miR196a in triplicate cultures, harvesting 24h after transfection. We transfected pre-miR196 in okf4 immorualised normal oral keratinocytes in duplicate culture. Successful transfection was ensured by qPCR. 16 Samples in total. 2 high expressing cells: control and anti-miR transfected, in triplicate. 1 low expressing cell: control and premiR transfected, in duplicate
Project description:Analysis of long-acting progestin-only contraceptive (LAPC)-mediated differential gene expression in vascular smooth muscle cells (VSMCs). We tested the hypothesis that LAPCs influence expression of several common genes associated with differentiation, survival and migration of VMSCs. Results provide important information of several common and unique gene expression profiles in cultured VSMC treated with ETO, M or P. Among ETO and M responsive genes, up- or down-regulated common genes were determined and used further confirmation and in vitro functional analyses. Total RNA (n=3) obtained from cultured aortic VSMCs treated with vehicle only (CON) or ETO or M or P for 6h.
Project description:Analysis of differential gene expression in Human endometrial endothelial cells (HEECs) incubated with CMS derived from human endoemtrial stromal cells (HESCs) treated by LAPCs. We tested the hypothesis that paracrine factors sectreted from HESCs treated LAPCs, etonogestrol (ETO) or medorxyprogesterone acetate (M) influence several common genes associated with survival in HEECs. Thefore, whole genome analyses were performed in HEECs treated with HESC derived CMS obtained from vehicle (estrodial (E) as control) or progesterone (P) or ETO or M incubations under hypoxia (HX) and normoxia (NX). Results provide important information of several common and unique gene expression profiles in cultured HEECs treated with HESC CMS from P or ETO or M incubations. Among M and ETO responsive genes, up- or down-regulated survival related common genes were determined and used further confirmation and in vitro functional analyses. Total RNA (n=3) obtained from cultured HEECs incubated 6h with vehicle- or P4- or ETO- or MPA- treated NX or HX conditioned HESC-derived CMS.
Project description:Expression profiles of normal colons in ApcMin mice that were fed methyl deficient diet for 11 weeks vs controls Expression profiles of normal colons in ApcMin mice that were fed methyl deficient diet for 11 weeks vs controls fed methyl donor sufficient diets
Project description:Presently RWA resistant genes are believed to be single dominant genes. However, some studies have shown that this may not necessarily be the case. It is possible that RWA resistance in wheat may be a function of multiple alleles on a single site, quantitative trait loci (QTL), pathway signalling differences or varying methylation patterns. This study aims to investigate the underlying genetic controls of RWA resistance. Thereby allowing breeders to make informed choices during selection phases.
Project description:Molecular mechanisms responsible for abnormal endometrial vasculature in women receiving long-acting progestin-only contraceptives (LAPCs) are unknown. We hypothesize that LAPCs impairs vascular smooth muscle cell (VSMC) and pericyte proliferation and/or migration producing thin-walled hyperdilated fragile microvessels prone to bleeding. Proliferating cell nuclear antigen (PCNA) and alpha smooth muscle actin (aSMA) double-immunohistochemistry assessed VSMC differentiation and proliferation in endometria from women pre- and postDepoProvera (Depo) treatment and from oophorectomized guinea pigs (OVX-GPs) treated with vehicle, estradiol (E2), medroxyprogesterone acetate (MPA) or E2+MPA pellets. After treating cultured VSMCs with MPA or etonogestrel (ETO), whole genome profiling, proliferation and migration assays were performed. Endometrial vasculature of Depo-administered women displayed reduced M-DM-.SMA immunoreactivity and fewer PCNA (+) nuclei among aSMA (+) cells (P<0.008). Microarray analysis of VSMCs identified several MPA and ETO-altered transcripts regulated by STAT1 signaling (p<2.22x10-6), including chemokine (C-C motif) ligand 2 (CCL2). Both MPA and ETO reduced VSMC proliferation and migration (p<0.001), recombinant CCL2 reversed this progestin inhibition, and, in turn, a STAT1 inhibitor abolished these CCL2 effects. Similarly, the endometria of MPA treated OVX-GPs displayed decreased aSMA staining and fewer PCNA (+) nuclei in VSMC (p<0.005). In conclusion, LAPCs promote abnormal endometrial vessel formation by inhibiting VSMC proliferation and migration. Total RNA (n=3) obtained from cultured TDCs incubated 6h with estradiol or estradio + medroxyprogesterone acetate with or without 1 ng/ml IL-1M-NM-2 for 6h.
Project description:We show here that the T helper-fate is not fixed and that mature antigen-stimulated CD4 T cells can switch off Thpok expression and reactivate CD8- lineage genes. This unexpected plasticity results in the post-thymic termination of the T helper- program and the functional differentiation of distinct MHC class II restricted CD4 cytotoxic T lymphocytes Small intestine CD4 intraepithelial T lymphocytes from ThPOK-GFP reposter mice were isolated and sorted (FACS Aria) based on ThPOK and CD8aa expression. Cell were isolated either from non-experimental ThPOK-GFP reporter mice (WT) or after transfering CD4 naive T cells from ThPOK-GFP reporter mice to RAG-/-recipient animals (TR) as an experimental colitis model. Experimet was done in duplicate.
Project description:Stomata are highly specialized organs which consist of pairs of guard cells and regulate gas and water vapor exchange in plants. While early stages of guard cell differentiation have been described and were interpreted in analogy to processes of cell type differentiation in animals, the downstream development of functional stomatal guard cells remains poorly understood. We have isolated an Arabidopsis mutant, scap1 (stomatal carpenter 1), that develops irregularly shaped guard cells and lacks the ability to control stomatal aperture, including CO2-induced stomatal closing and light-induced stomatal opening. SCAP1 was identified as a plant-specific Dof-type transcription factor expressed in maturing guard cells but not in guard mother cells. SCAP1 regulates the expression of genes encoding key elements of stomatal functioning and morphogenesis, such as a K+ channel protein, MYB60 transcription factor, and pectin methyl esterase. Consequently, ion homeostasis was disturbed in scap1 guard cells, and esterification of extracellular pectins was impaired so that the cell walls lining the pores did not mature normally. We conclude that SCAP1 regulates essential processes of stomatal guard cell maturation and functions as a key transcription factor regulating the final stages of guard cell differentiation. We isolated guard cell protoplasts from 4-week-old WT(Col-0) and scap1 mutant plants and extracted RNA independently. Four biological replicates were performed for each experiment.