Interferon stimulation creates chromatin marks and establishes transcriptional memory | PNAS
BM-derived macrophages were cultured in DMEM/F12 medium containing 10% FBS, 20% L929-conditioned medium as a source of ... in serum-free and antibiotics-free medium for 1 h at 37 °C, washed, and allowed to proceed in culture for 24 h. Cells were ... However, H3.3 siRNA, when added to naïve cells, did not inhibit ISG induction as much under these conditions. These data are in ... For viral titration, plaque assays were performed with L929 cells incubated with serial dilutions of culture supernatants (10−2 ...
Gleevec shifts APP processing from a β-cleavage to a nonamyloidogenic cleavage | PNAS
Secreted Aβ 40 levels from culture medium were measured by sandwich ELISA. n = 3. *P , 0.05; **P , 0.01, Students t test. (D) ... In contrast, we demonstrated that the Gleevec-induced APP-CTFs were greatly reduced under these conditions. Furthermore, ... Human Aβ 40/42 was measured in cell culture media and homogenized mouse brain extracts using Aβ 40/42 sandwich ELISA kits ( ... stock solutions (10 and 20 mM) were prepared in DMSO (Sigma-Aldrich). Each Inhibitor was rapidly mixed in culture medium (1: ...
Aerobic bacterial methane synthesis | PNAS
All strains were cultured in SAR media containing NH4+, MeA, or glycine betaine as the sole nitrogen source (treatment symbols ... 3B). Cultures initiated at cell densities equivalent to mid log phase under these growth conditions (starting optical density ... A 100 mL culture of E. coli BL21 was grown to stationary phase in M9 minimal media (84), collected by centrifugation, suspended ... For culture characterizations, Acidovorax was grown in liquid SAR-MeA media containing glucose as the sole carbon source (4 g/L ...
Buffering of deoxyribonucleotide pool homeostasis by threonine metabolism | PNAS
Twenty milliliters of culture was collected by vacuum filtration and immediately washed with ice-cold medium, and filters were ... AUGC is plotted on the y axis for each culture/condition. Error bars represent standard deviation. ... Strains were grown overnight in liquid medium at 30°C to a concentration of ≈3 × 106 cells/ml and diluted to prewarmed medium ... Media supplementation with amino acids was used to test whether uptake of extracellular threonine was able to suppress the ...
Nonoverlapping roles of PD-1 and FoxP3 in maintaining immune tolerance in a novel autoimmune pancreatitis mouse model | PNAS
Under such conditions, self-reactive T cells may develop into the effector population. This could be an additive, Treg- ... The tissue was cut into about 3-5-mm-thick pieces and incubated 30 min in RPMI medium 1640 (Life Technologies, serum free) with ... After 10 d of culture, each group of cells was sorted again, and then 1 × 106 CD8+, ex-FoxP3, or the combination of CD8+ and ex ... FoxP3+ Treg cells constitutively express PD-1 under diverse conditions, in both mice and humans. In fact, Treg cells isolated ...
A ratiometric lectin microarray approach to analysis of the dynamic mammalian glycome | PNAS
The HL-60 cells were purchased from American Type Culture Collection and cultured in RPMI medium 1640 supplemented with 20% ... under fixed scanning conditions with a range of labeled glycoproteins. Therefore, only signals above the 1,000-A.U. threshold ... CHO Cell Culture.. We purchased Pro-5, Lec1, Lec2, and Lec8 CHO cell lines from American Type Culture Collection (Manassas, VA ... We cultured Pro-5 and Lec8 cell in tissue culture-treated, black-wall, clear-bottom 96-well plates (Greiner Bio-One). We ...
Elucidation of tonic and activated B-cell receptor signaling in Burkitt's lymphoma provides insights into regulation of cell...
Cell Culture, BCR Stimulation, and Cell Lysis.. All cell lines were cultured in RPMI medium (Invitrogen) supplemented with 10- ... 2 B and C). An shRNA induction duration of 18 h was identified as the optimum condition for the analysis of tonic BCR signaling ... DG75 cells were cultured in SILAC medium and treated as indicated. Lysates were processed as described in A. (C) DG75 and Daudi ... DG75 cells were cultured in SILAC medium as indicated and were left untreated, or were BCR-stimulated, for 2, 5, 10, or 20 min ...
Critical role for Epac1 in inflammatory pain controlled by GRK2-mediated phosphorylation of Epac1 | PNAS
Cell Culture and Transfections.. HEK293 and N2A cells were cultured in high-glucose Dulbeccos modified Eagle medium (GE ... 6D). Under baseline conditions, the stimulus/response curve was similar in HEK and HEK-GRK2 cells expressing Piezo2 and Epac1. ... Chronic pain is associated with multiple inflammatory conditions, such as rheumatoid arthritis, inflammatory bowel disease, ... 50 cells per condition. (Scale bars, 25 µm.) *P , 0.05. (C) Rap1 activation in N2A cells transfected with WT Epac1 (ICUE1-WT) ...
Antibiotic treatment enhances the genome-wide mutation rate of target cells | PNAS
... cells were cultured on a tissue culture rotator for about 17 h to OD650 ∼ 1. Culture was then serially diluted and about 1,500 ... Strains, Media, and Antibiotics.. All wild-type (laboratory designation name, PFM2), ∆mutY (PFM6), ∆mutS (PFM342), and marR:M1V ... This discrepancy may be a result of the cells growing in different physiological conditions in the experiment of Kohanski et al ... Cells were cultured overnight (∼17 h) and then serially diluted. About 1,500 cells were then plated onto LB plates containing 0 ...
A soluble bone morphogenetic protein type IA receptor increases bone mass and bone strength | PNAS
Conditioned medium containing mBMPR1A-mFc was purified using two-step column chromatography, dialyzed into PBS, and purity ... RNA was isolated, using UltraSpec reagent (Biotecx), from SaOS2 cells cultured for 48 h with or without BMP2 (25 ng/mL; R&D ... Supernatant were isolated from SaOS2 cells cultured for 72 h with or without BMP2 and in the presence or absence of mBMPR1A-mFc ... 100 kDa under nonreducing conditions (Fig. S1A). SDS/PAGE and size exclusion chromatography showed that mBMPR1A-mFc was ∼95% ...
The role of worldviews in the governance of sustainable mobility | PNAS
"Cars and culture in Munich and Birmingham: The case for cultural pluralism" in Politics, Policy, and Culture, D. J. Coyle, R. ... The ways of life compete with but ultimately, depend on each other-the requisite variety condition set out in Cultural Theory- ... and censorship of media, which portray the hierarchical faith in social conformity. Hence, we labeled it as hierarchy. The ... Therefore, environmental concern is rooted in an egalitarian culture (20).. By contrast, a hierarchical culture is ...
Farming and public goods production in Caenorhabditis elegans populations | PNAS
C. elegans Strains and Culture.. N2 Bristol (laboratory wild type) and AT10 (srf-3 (yj10)) (mutant type) were obtained from the ... Worms disperse bacteria in natural settings such as rotting fruit (E) and soil-like porous medium (F). Arrows indicate initial ... under identical initial conditions, the N2 population grew larger than that of the srf-3 worms (Fig. 2C; brood sizes of both ... If there is no cost to farming, farmers and nonfarmers perform equally well in mixed cultures, consistent with our experimental ...
Ancient Maya wetland fields revealed under tropical forest canopy from laser scanning and multiproxy evidence | PNAS
The collective area of wetland complexes is 14.08 km2 within one medium-sized watershed with a dry season discharge of ∼6-7 m3· ... This represents a fortunate set of conditions given that all of the previous archaeological lidar research in the region has ... Wetland agriculture was crucial to ancient cultures, but no previous study coupled lidar with multiproxy evidence to ... during regionally dry conditions, may have reattracted or maintained less intensive land uses like hunting, fishing, gathering ...