dnase  (Worthington Biochemical)

Journal: Molecular and Cellular Biology

Article Title: Epigenetic Control of Cell Cycle-Dependent Histone Gene Expression Is a Principal Component of the Abbreviated Pluripotent Cell Cycle

doi: 10.1128/MCB.00736-12

Figure Lengend Snippet: Chromatin architecture of the human histone H4 gene in hES cells. (A) DNase I hypersensitivity at the human histone H4 gene ( HIST2H4 ). Nuclei were isolated from hES cells (lanes 4 to 6) and normal diploid fibroblasts (TIG-1 and IMR-90; lanes 7 to 9 and

Article Snippet: Nuclei were then resuspended in RSB buffer supplemented with 1 mM CaCl2 and incubated with increasing concentrations of DNase I (DPRF; Worthington Biochemical Corporation, Lakewood, NJ) for 10 min at room temperature with gentle agitation.

Techniques: Isolation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: STAT3, STAT4, NFATc1, and CTCF regulate PD-1 through multiple novel regulatory regions in murine T cells

doi: 10.4049/jimmunol.1302750

Figure Lengend Snippet: The Pdcd1 locus contains multiple inducible DNase I hypersensitive sites

Article Snippet: Concentration ranges for DNase I were determined empirically for each lot and cell type by titrating DNase I for its ability to digest CR-C as the positive control but not regions previously found to be resistant to DNase I (e.g., +6.3 region).

Techniques:

Journal: Microbes and infection / Institut Pasteur

Article Title: Extracellular traps are associated with human and mouse neutrophil and macrophage mediated killing of larval Strongyloides stercoralis

doi: 10.1016/j.micinf.2014.02.012

Figure Lengend Snippet: NETs are required for larval killing by human neutrophils in vitro. A) Human macrophages (Mϕ) and neutrophils (PMN) cultured in vitro with 50 larvae (L3) for 48 h. The cultures were treated with 100 U/ml of DNase I to block NET formation and assessed

Article Snippet: Treatment with DNase I eliminated the presence of released DNA, but did not block killing of the larvae by mouse neutrophils and macrophages in vitro.

Techniques: In Vitro, Cell Culture, Blocking Assay

Journal: The EMBO Journal

Article Title: A cell-cell signaling peptide activates the PlcR virulence regulon in bacteria of the Bacillus cereus group

doi: 10.1093/emboj/cdf450

Figure Lengend Snippet: Fig. 8. ( A ) β-galactosidase activity of the B.thuringiensis 407 Cry – A ′ Z Δ papR mutant strain. The cells were grown at 37°C in LB medium and each peptide (OS5, OS5-I1, OS5-M1 and OS5-V1; all 0.5 µM) was added at t 1 (1 h after the onset of the stationary phase). The amino acid sequences of the peptides are as follows: OS5, LPFEF; OS5-I1, IPFEF; OS5-M1, MPFEF; OS5-V1, VPFEF. ( B ) DNase I footprinting analysis of PlcR binding to the plcA promoter region in the presence of various pentapeptides. Lane 1, G+A Maxam and Gilbert reaction; lane 2, no protein; lane 3, 1.2 µg of PlcR (35 pmol); lane 4, 1.2 µg of PlcR and of OS5 10 µg; lane 5, 1.2 µg of PlcR and 10 µg of OS5-I1; lane 6, 1.2 µg of PlcR and 10 µg of OS5-M1; lane 7, 1.2 µg of PlcR and 10 µg of OS5-V1. The region protected by PlcR is indicated by a bracket and corresponds to the PlcR box upstream from the plcA gene.

Article Snippet: Reactions were performed in a 20 µl volume, and concentrations of MgCl2 and CaCl2 were adjusted to 5 and 0.5 mM, respectively, before adding 5 ng of DNase I (Worthington Biochemical).

Techniques: Activity Assay, Mutagenesis, Footprinting, Binding Assay

Journal: The EMBO Journal

Article Title: A cell-cell signaling peptide activates the PlcR virulence regulon in bacteria of the Bacillus cereus group

doi: 10.1093/emboj/cdf450

Figure Lengend Snippet: Fig. 6. DNase I footprinting analysis of PlcR binding to the plcA promoter region. Radiolabeled fragments (50 000 c.p.m.) were incubated with various concentrations of PlcR and OS5. ( A ) Lane 1, G+A Maxam and Gilbert reaction; lane 2, no protein; lane 3, 0.1 µg of PlcR; lane 4, 10 µg of PlcR; lane 5, 20 µg of OS5; lane 6, 0.1 µg of PlcR and 10 µg of OS5; lane 7, 0.1 µg of PlcR and 20 µg of OS5; lane 8, 10 µg of PlcR and 10 µg of OS5; lane 9, 10 µg of PlcR and 20 µg of OS5. The region protected by PlcR is indicated by a bracket. ( B ) Lane 1, G+A Maxam and Gilbert reaction; lanes 2–8, 50 pmol of PlcR (1.7 µg) and OS5: lane 2, 1000 pmol (0.65 µg); lane 3, 500 pmol; lane 4, 150 pmol; lane 5, 70 pmol; lane 6, 35 pmol; lane 7, 10 pmol; lane 8, 5 pmol; lane 9, 50 pmol of PlcR; lane 10, neither protein nor peptide. ( C ) plcA promoter region. The DNase I-protected area is indicated by a bracket and the PlcR box is in bold. Positions are relative to the transcription start point. The –35 and –10 promoter regions, and the transcription start (+ 1) of the plcA ).

Article Snippet: Reactions were performed in a 20 µl volume, and concentrations of MgCl2 and CaCl2 were adjusted to 5 and 0.5 mM, respectively, before adding 5 ng of DNase I (Worthington Biochemical).

Techniques: Footprinting, Binding Assay, Incubation