Sentences with phrase «ng -rcb-»

For a complete list of authors and funding agencies, see Nature Genetics (DOI: 10.1038 / ng.888).

OptiMat was made by mixing equal volumes of 666 ng / µl Fibronectin, 337 ng / µl Collagen I and 1 µg / µl Collagen IV.

Cells were cultured on irradiated mouse embryonic fibroblasts (MEF) in DMEM / F12 culture medium supplemented with KnockOut serum replacer (20 %), non-essential amino acids (0.1 mM), L - glutamine (1 mM), ß - mercaptoethanol (0.1 mM) and 100 ng / ml zebrafish basic fibroblast growth factor.

DNA derived from FFPE (20 ng) was amplified by PCR using AmpliSeq primers and HiFi master mix (Ion AmpliSeq v. 2.0).

(B) The expression of SCP1 and SCP3 in monkey testis (5 years old), ES cells (ES), and developing EBs (days 14, 21, and 28) in the presence or absence of BMP4 (100 ng / ml), RA (1 µM), and SCF (100 ng / ml) was examined using RT - PCR.

Lanes 1 and 20, 100 - bp ladder; lanes 2 - 15 are results from CFS patients; lanes 16 - 18 assay sensitivity controls consisting of 10, 3 and 1 copies of XMRV VP62 plasmid DNA diluted in a background of 250 ng of human PBMC DNA; lane 19, water control.

Using a cutoff level of 6 ng / L, offered as optimal by the authors, the proportion meeting the criterion increased to 39 % and the NPV was 99.0 % (CI, 97.5 % -99.7 %).

Firstly, SH - SY5Y cells where treated with 200 ng / ml of GDF5 for various time points for up to 60 min to examine Smad1 / 5 phosphorylation, or for 72 h to examine neurite growth.

(B) Neurite length of GDF5 - treated (200 ng / ml daily for 72 h) SH - SY5Y cells at 72 h. (C — F) p - Smad1 / 5 levels as determined by (C, D) Western blots or (E, F) immunocytochemistry in SH - SY5Y cells 24 h after transfection with control miRNA or miR - 181a inhibitor.

Where indicated, cells were treated daily with 10 or 200 ng / ml of growth differentiation factor (GDF) 5 (GDF5)(Biopharm GmbH), which are maximal saturating concentrations in primary and SH - SY5Y cell cultures respectively [17,41].

Data using a single sample with a cutoff at 2 ng / L were not reported.

Subconfluent HUVEC cultures were stimulated with PMA (50 ng / mL) for 4 h in the absence (controls) or presence of different concentrations of kahweol.

PCR was performed on 500 ng of genomic DNA using specific primers against MSTN, BLG, PrP and NUP (Table S1).

Sustained hyperleptinemia of 8 ng / ml was induced for 28 days in normal Wistar rats by infusing a recombinant adenovirus containing the rat leptin cDNA (AdCMV - leptin).

The mean Cmax was 114 ng / ml occurring at 1 hour after dosing, and the area under the serum concentration versus time curve was 573 ng × h / ml, with a terminal phase half - life of 1.9 hours observed.

(A, B) Neurite length of neurons in E14 rat VM cultures transfected with control siRNA, Smad4 siRNA or Smad1 siRNA or Smad5 siRNA and a GFP - expressing plasmid before being cultured with or without 10 ng / ml GDF5 for 72 h (*** P < 0.001 compared with control; ANOVA with post-hoc Tukey's test; 40 cells for each group per experiment; number of repetitions (n) = 3 experiments).

MNK - 3 cell line [45] and clone B3 were maintained in DMEM (Corning; Tewksbury, MA) with 10 % heat - inactivated FBS (Gemini Bio-Products; West Sacramento, CA), 2 mM GlutaGro (Corning), 1 mM sodium pyruvate (GE Healthcare HyClone; Logan, UT), 55 μM β - mercaptoethanol (Sigma), 10 mM HEPES (Corning), 50 μg / ml gentamycin (Amresco; Solon, OH), 100 U / ml penicillin (Gemini Bio-Products), 100 U / ml streptomycin (Gemini Bio-Products) and 10 ng / ml recombinant mouse IL - 7 and IL - 15 (eBioscience; San Diego, CA or Peprotech; Rocky Hill, NJ).

PCR product was purified using Chromaspin technology (Clontech): 50 — 100 ng of PCR product was used for sequencing with the DYEnamic ET Dye Terminator Kit (Amersham Biosciences) according to the manufacturer's instructions.

Dual transfection was carried out using 12 μl of polyethylenimine to 3 μg of NFκB luciferase construct and 60 ng of Renilla luciferase construct.

This input DNA represents about 350 ng of PBMC DNA which is similar to the amount used by others [11 — 13, 15, 16], thus not affecting the sensitivity of our results.

Human tonsillar lymphocytes were treated with lethal toxin or E687C mutant lethal toxin (1.0 μg / ml) for 3 hr followed by IL - 23 (50 ng / ml) stimulation for 18 hr.

For standard PCR, 500 ng of DNA from each tissue was added to the amplification.

This technique utilized 250 ng of RNA for the first step, which may have been too little given the size of monkeys and volume of tissue.

Cells were restimulated for 5 hr in the presence of BFA with or without 5 μg / ml phorbol 12 - myristate 13 - acetate (PMA)(Sigma), 0.5 μg / ml ionomycin (Sigma) and 50 ng / ml IL - 23.

The conventional hESC medium contained 80 % (vol / vol) KnockOut Dulbecco's Modified Eagle's Medium (DMEM) and 20 % (vol / vol) KnockOut serum replacement supplemented with 2 mM Glutamax, 0.1 mM β - mercaptoethanol, 0.1 mM MEM non-essential amino acids (Cambrex Bio Science, Walkersville, Inc., Walkersville, MD), 50 U / ml penicillin - 50 μg / ml streptomycin (Cambrex Bio Science) and 8 ng / ml recombinant human fibroblast growth factor (bFGF, R&D Systems, Minneapolis, MN).

Treatment with 10 ng / ml of GDF5 for 72 h led to significant increases in neurite length (P < 0.001)(Figure 2A, C).

(A) VASA expression in EBs cultured in the presence or absence of BMP4 (100 ng / ml), RA (1 µM), and SCF (100 ng / ml) was compared using quantitative RT - PCR.

Oligodendrocyte differentiation was induced by culturing the cells in DMEM / F12, 1 % N2, and IGF - 1 (200 ng / ml).

Cells were cultured in RPMI with 10 % FBS, 100 U / ml penicillin, 100 U / ml streptomycin, 1 mM sodium pyruvate, non-essential amino acids, 50 μM β - mercaptoethanol, 2mM glutamine and recombinant human IL - 2 (20 ng / ml), IL - 7 (20 ng / ml), SCF (20 ng / ml), FLT3L (10 ng / ml), IL - 15 (10 ng / ml) for up to three weeks.

Single - cell suspensions were obtained from spleen, mesenteric lymph node (MLN), and organs of mice and were stimulated in vitro with 50 ng / ml PMA and 500 ng / ml ionomycin (both from Sigma - Aldrich) for 5 — 6 h in the presence of GolgiStop or GolgiPlug (BD Biosciences).

Human tonsillar lymphocytes were treated with increasing concentrations (0.01 — 10 μg / ml) lethal toxin for 3 hrs followed by IL - 23 (50 ng / ml) stimulation for 18 hr.

The precipitated cells were resuspended in the serum - free medium containing DMEM / F12 (Invitrogen), B27 (2 %)(Invitrogen), bFGF (20 ng / mL)(Invitrogen) and EGF (20 ng / mL)(R&D), and plated onto a 60 mm low - attachment dish.

RAW264.7 medium was replaced by conditioned medium harvested from tumor cells and supplemented with recombinant murine sRANKL (50 ng / ml).

Differentiation of hGPCs was induced at 2500 cells / cm2 by replacing bFGF with either 20 ng / ml BMP - 4 (R&D) or 10 ng / ml CNTF (Peprotech) and allowed to differentiate for 7 days prior to harvest.

MNK - 3 cells (an ILC3 cell line) were cultured in MNK - 3 media (DMEM (High glucose), 10 % FBS, 100 U / ml penicillin, 100 U / ml streptomycin, 2 mM Glutagro, 1 mM sodium pyruvate, 10 mM HEPES, 55 μM β - mercaptoethanol, 5 μg / ml gentamicin, 10 ng / ml IL - 7 and 10 ng / ml IL - 15).

(A-C) Rag1 - / - splenocytes were pretreated with (A) either lethal toxin (LeTx, lethal factor + protective antigen) or lethal factor (LF)(1 μg / ml) or (B) increasing doses of lethal toxin or (C) with enzymatic mutant toxin (E687C) for 3 hr followed by IL - 23 (50 ng / ml) stimulation for 18 hr.

and cultured in 5 % O2 / 7 % CO2 in Bottenstein - Sato F12 medium with 10 ng / ml human recombinant basic fibroblast growth factor (Peprotech) on a substrate of 1 µg / cm2 fibronectin (Chemicon) and 0.5 µg / cm2 laminin (Invitrogen).

(A) Human tonsillar lymphocytes or (B) sorted Lin - CD127 + cells were pretreated with lethal toxin (1 μg / ml) for 2 — 3 hr followed by IL - 23 (50 ng / ml) stimulation for 18 hrs.

300,000 cells were plated per well of a 24 - well plate (minimum 10 wells) and mixed with virus (80 ng p24 / 106 cells) diluted in 100 µl of fresh medium.

After ten weeks in culture, 10 ng / ml leukemia inhibitory factor (Chemicon, Temecula, CA) was also added.

For lethal toxin experiments MNK - 3 cells were cultured in media containing 10 ng / ml IL - 7.

100,000 — 500,000 splenocytes per well were incubated in a round bottom 96 well plate in IMDM media supplemented with 10 % FBS, 100 U / ml pencillin, 100 U / ml streptomycin, IL - 2 (20 ng / ml) and IL - 7 (10 ng / ml).

Cells after toxin treatment were stimulated with recombinant mouse or human IL - 23 (50 ng / ml)(eBioscience) for 18 hrs.

The findings were published this week in Miotto et al, Multiple populations of artemisinin - resistant Plasmodium falciparum in Cambodia, Nature Genetics, 2013; Jun; 45 (6): 648 - 55; advance online publication, 28 April 2013 (doi: 10.1038 / ng.2426).

The probability of having a newly diagnosed or known diabetes more than doubled in women with levels below a cut - off of 15 ng / mL.

PC12 cells were plated in growth conditioned medium (CM) prepared from HT22 cells incubated overnight plus or minus 100 nM J147, and as controls fresh DMEM plus 100 nM J147 or fresh DMEM plus 50 ng / ml NGF.

Genomic DNA was isolated using the QIAamp DNA Blood Mini Kit (Qiagen, Valencia, CA, USA), quantified by the Qubit 2.0 fluorometer (Life Technologies, Thermo Fisher Scientific, Waltham, MA, USA), assessed on the Agilent TapeStation 2,200 (Agilent Technologies Inc., USA) and ∼ 120 ng was subsequently subjected to whole exome DNA library construction using the Ion AmpliSeq Whole Exome RDY (Life Technologies, Thermo Fisher Scientific, Waltham, MA, USA) essentially as described in the manufacturer's protocol, with barcode incorporation.

Quantification was performed in comparison to a RNA - standard of 20 ng / ml -1000 ng / ml RNA supplied with the kit and a standard curve was established.

Each assay (15 µL total volue) contained nuclease free water (5 - 10 µL), cDNA template (2 ng / µL), and gene - specific primers (4.5 pm / µL).

In the meantime, 1,000 ng ml − 1 VEGFa (RELIATech, # 300 - 036) and FGF2 (RELIATech, # 300 - 003L) was added to the methocel / fibrinogen / EC basal medium mix.