1,775,887 research outputs found
GAL-Repository/EDA_Stuff: v1.0.2
No full textFull Changelog: https://github.com/GAL-Repository/EDA_Stuff/commits/v1.0.
Activation of Silent gal Genes in the lac-gal Regulon of Streptococcus thermophilus
Get PDFStreptococcus thermophilus strain CNRZ 302 is unable to ferment galactose, neither that generated intracellularly by lactose hydrolysis nor the free sugar. Nevertheless, sequence analysis and complementation studies with Escherichia coli demonstrated that strain CNRZ 302 contained structurally intact genes for the Leloir pathway enzymes. These were organized into an operon in the order galKTE, which was preceded by a divergently transcribed regulator gene, galR, and followed by a galM gene and the lactose operon lacSZ. Results of Northern blot analysis showed that the structural gal genes were transcribed weakly, and only in medium containing lactose, by strain CNRZ 302. However, in a spontaneous galactose-fermenting mutant, designated NZ302G, the galKTE genes were well expressed in cells grown on lactose or galactose. In both CNRZ 302 and the Gal+ mutant NZ302G, the transcription of the galR gene was induced by growth on lactose. Disruption of galR indicated that it functioned as a transcriptional activator of both the gal and lac operons while negatively regulating its own expression. Sequence analysis of the gal promoter regions of NZ302G and nine other independently isolated Gal+ mutants of CNRZ 302 revealed mutations at three positions in the galK promoter region, which included substitutions at positions -9 and -15 as well as a single-base-pair insertion at position -37 with respect to the main transcription initiation point. Galactokinase activity measurements and analysis of gusA reporter gene fusions in strains containing the mutated promoters suggested that they were gal promoter-up mutations. We propose that poor expression of the gal genes in the galactose-negative S. thermophilus CNRZ 302 is caused by naturally occurring mutations in the galK promoter.
Gal-9 epitope recognition by Gal-Nab1 and Gal-Nab2.
No full textA. Apoptotic cell death assessed by flow cytometry in CD3+ T cells treated for 36h with gal-9 (gal-9S; 40 nM) alone or in combination with mAbs (ctrl IgG1, Gal-Nab1, Gal-Nab2; 67 nM) pre-incubated or not with scramble peptide (Scr.) or gal-9 CTB-peptide (6.7 μM). B. Human recombinant gal-9 was immobilized in 96-wells plates and binding of Gal-Nab1 and Gal-Nab2 mAbs was measured after pre-incubation with overlapping peptides representative of human gal-9, as described under “Material and Methods”. Gal-Nab1 (left) or Gal-Nab2 (right) were then detected using secondary HRP-conjugated anti-mouse antibodies. Percentages of inhibition induced by each peptide were calculated from the absorbance data as described under “Material and Methods”.</p
GAL-Repository/SARS-CoV-2_BW.1: v1.0.1
No full textFull Changelog: https://github.com/GAL-Repository/SARS-CoV-2_BW.
Anti-gal-9 mAbs efficiently neutralize gal-9-induced apoptosis in primary T cells.
No full textCD3+ T-cells were isolated from healthy donors, activated by a combination of CD3/CD28 antibodies and treated or not with gal-9 (gal-9S; 40 nM) alone or in combination with lactose (5 mM), control isotype mAbs (ctrl IgG1) or anti-gal-9 mAbs (Gal-Nab1 and Gal-Nab2) at 67 nM (i.e. 10 μg/mL). After 36 h, they were subjected to annexin-V/PI staining and flow cytometry analysis. A. Examples of flow cytometry plots for purified CD3+ cells from one donor. B. Synthesis of data from 3 similar experiments made with CD3+ cells from 3 donors. C and D. Dose-response curves for apoptotic cell death (annexin-V+ PI+) (C) or PS translocation (annexin-V+ PI-) (D) in activated CD3+ cells treated for 36h with gal-9 combined with increasing concentrations of Gal-Nab1 and Gal-Nab2 (0.3 to 100 nM). Empty squares indicate the percentages obtained in conditions without gal-9. Black crosses indicate the percentages obtained with isotype control IgG1 mAbs used at maximal concentration (100 nM). Data are presented as means ± SEM of three independent experiments made with three distinct donors.</p
Gal-3 expression in microglia cells.
No full textImmunohistochemical staining of Gal-3 (red) expressing microglia. All microglia were detected using the microglia marker Iba1 (green). At 0 DIV no Gal-3-expressing cells were found (A, B). In controls, at 3, 4 and 7 DIV Iba1/Gal-3 co-expressing cells were found and only in the GCL (C, D, G, H, K, and L). LPS-treated retinas displayed larger numbers of Iba1/Gal-3 co-expressing cells that were located in the GCL, INL and OPL at 3, 4 and 7 DIV. Scale bar: 200 μm.</p
Gal-9 binds and recruits to <i>Mtb</i>.
No full text(A) Experimental design for Mtb pull-down mass spectrometry identification of Mtb-binding proteins. (B) Domain organization of Gal-9. CRD, carbohydrate recognition domain. (C) Immunoblot of in vitro binding reactions between indicated pathogens and Gal-9-FLAG THP-1 lysate, probed with anti-FLAG antibody; IN, input; Lm, Listeria monocytogenes; Stm, Salmonella enteria serovar Typhimurium; Cn, Cryptococcus neoformans; Mtb, Mycobacterium tuberculosis. (D) Confocal microscopy of WT BMMs infected with WT or ΔeccC Mtb-GFP (MOI = 2) 8 hours post-infection and immunostained for endogenous Gal-9 and Gal-3. (E) Quantification of Mtb-GFP colocalization with Gal-9 or Gal-3 at indicated time points. Figures represent two independent experiments (D, E). An average of 882 cells were analyzed per technical replicate (D, E). Error bars represent SD from 3 technical replicates. The schematic was created with BioRender.com.</p
X-gal-negative Newly Formed Cardiomyocytes Increase Following Cardiac Injury.
No full text(A) X-gal staining and immunofluorescent images of sections comparing sham and myocardial infarction (MI) CreLacZ mice: left panel, sham; middle panel, MI remote area; and right panel, MI area. The upper panels show X-gal-stained images. The bottom panels show the corresponding immunofluorescent images (SA-actinin, green; laminin, red). Asterisks show X-gal-negative cardiomyocytes. Scale bar, 20 μm. The X-gal-negative cardiomyocytes are further enlarged in the inset. Scale bar, 10 μm. (B) Number of X-gal-negative cardiomyocytes per area 3 months after MI: sham (n = 5) and MI (n = 5). *p < 0.05. (C) Comparison of the number of X-gal-negative cardiomyocytes in the MI remote and MI area 3 months after MI (n = 5 per area). *p < 0.05. (D) Left: comparison of the cross-sectional area between X-gal-negative (MI2w neg) and -positive (MI2w pos) cardiomyocytes at 2 weeks after MI. Right: comparison of the cross-sectional area of X-gal-negative cardiomyocytes at 2 weeks (MI2w neg) and 6 months (MI6m neg) after MI. n = 50–66 cardiomyocytes pooled from two MI mice per group. *p < 0.05. The Mann–Whitney U-test was used for statistical analysis.</p
H-gal-GP is flexible.
No full textThe three main ways in which H-gal-GP appears to move, as determined by principal component analysis of multi-body refinement, are illustrated in each row A-C.</p
Subunit fitting to H-gal-GP EM density.
No full textA EM density of one-winged H-gal-GP (4.5 Å map) fitted with models of aspartyl protease PEP 1 (red), MEP3 (dark blue) and cysteine protease (cyan). A cropped view of A, through the centre of the H-gal-GP complex (B) and cropped to just show two of the MEP3 domains (C). D H-gal-GP map viewed from the base showing the fitting of the MEP subunits. Fitted subunits of the H-gal-GP complex viewed from the side (E) and base (F) and colored red (PEP1), cyan (cysteine protease) and orange, blue, green and grey for the four MEPs.</p
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