Brain urea increase is an early Huntington's disease pathogenic event observed in a prodromal transgenic sheep model and HD...
1994) Cloning and functional expression of a urea transporter from human bone marrow cells. J Biol Chem 269:31649-31652. ... 1997) Cloning and characterization of the urea transporter UT3: Localization in rat kidney and testis. J Clin Invest 99:1506- ... As cell dysfunction is initiated before onset of symptoms it is likely that an effective therapy would need to be applied early ... To reduce the likelihood of cell death as a cause for the phenotype, urea levels were quantified specifically in brain tissue ...
Aerobic bacterial methane synthesis | PNAS
Positive clones were DNA prepped and transformed into BL21 (DE3) cells (Novagen). ... Cells were incubated at 25 °C for 8 h, then spun down (3,000 rcf, 20 min, 4 °C). The resulting cell pellet was stored at −80 °C ... The 16S rRNA genes were PCR cloned using 27F and 1492R PCR primers (SI Appendix, Table S2). Amplicons were individually cloned ... Transposon Mutagenesis, plp-aat Cloning, and Complementation.. Methods for transposon mutagenesis, genome walking, and cloning ...
Critical role for Epac1 in inflammatory pain controlled by GRK2-mediated phosphorylation of Epac1 | PNAS
The two right lanes represent two different GRK2-overexpressing clones. (D) Rap1 activation in N2A-GRK2 or -K220R cells ... 22-26 cells) or control HEK-PE cells (n = 20-22 cells). The 8-pCPT or bath solution was added, and cells were voltage-clamped ... B) Rap1 activation in HEK293 (HEK) or HEK293 cells overexpressing GRK2 (HEK-GRK2) cells. Cells transfected with HA-Epac1 were ... A) Pull‐down of GRK2 with Epac1 in N2A cells. YFP‐Epac1 or GFP were co‐expressed with GRK2‐V5 in N2A cells. Lysates were ...
A soluble bone morphogenetic protein type IA receptor increases bone mass and bone strength | PNAS
Human SaOS-2 cells (a human osteosarcoma-derived osteoblast cell line) were treated for 20 min with BMP2 and/or mBMPR1A-mFc ( ... The extracellular domain of murine BMPR1A was cloned into pAID4 to produce the mBMPR1A-mFc construct (Fig. 1A). The construct ... D) Cell-based analysis of the ability of mBMPR1A-mFc to inhibit BMP2 and BMP4-induced signaling. Experimental sample values are ... 2002) The Wnt antagonist Dickkopf-1 is regulated by Bmp signaling and c-Jun and modulates programmed cell death. EMBO J 21:966- ...
Profile of Charles M. Rice | PNAS
"Sangeetas cell culture system helped the cells retain their liver-like phenotype outside the liver," Rice says. Lofty yet ... "When we cloned and sequenced the viral RNA, it became clear that the yellow fever virus was remarkably different from Sindbis ... a technique that allows them to airlift single infected cells from a carpet of cells growing on dishes, to perform detailed ... Liver cells lacking any of the three proteins failed to take up the virus (7). "The finding stemmed from our efforts to develop ...
Buffering of deoxyribonucleotide pool homeostasis by threonine metabolism | PNAS
... median cell size increased whereas the relative number of cells and total cell volume (median cell size × median cell volume) ... pJH023 was constructed from pNEB193 (New England Biolabs, Ipswich, MA). The multiple-cloning site was expanded by annealing ... c) Relative increases in cell number (CNI), median cell size (CSI), and total cell volume (TCVI), for cultures shown in b. ... The total cell volume of each culture (median cell size × total cell number) was used for calculating intracellular dNTP pool ...
Immunization with a functional protein complex required for erythrocyte invasion protects against lethal malaria | PNAS
... live cells as measured by counting trypan blue-stained cells. 2 × 106 purified T cells were injected on days −1, 0, and +1, ... IgG Against the Pf3D7-AMA1-RON2L Complex Does Not Block Invasion by the Heterologous PfFVO Clone.. As discussed above, the ... These mice were challenged with 105 PyYM iRBCs on day 0. T cells from immunized animals were purified using a mouse pan T-cell ... but T cells were not (Fig. 1 H and I and Fig. S1C). Our data do not rule out the possibility of a role for T cells together ...
Nonoverlapping roles of PD-1 and FoxP3 in maintaining immune tolerance in a novel autoimmune pancreatitis mouse model | PNAS
Unlike Treg cells, the ex-FoxP3 cells are pathogenic cells with an effector/memory phenotype. Indeed, ex-FoxP3 cells were shown ... The antibodies conjugated with various fluorophores for flow cytometry included CD4 (clone: GK1.5), CD8 (53-6.7), CD25 (PC61.5 ... ex-FoxP3 T cells in autoimmune FoxP3-GFPcreKI/Y PD-1 KO mice. (A) FACS analysis of Tregs, ex-FoxP3 T cells, and CD25+ T cells ... B) Percentage of GFP−RFP+ (ex-FoxP3) T cells within RFP+ T cells (Treg plus ex-FoxP3) of CD4+ T cells or CD4−CD8+ thymocytes ( ...
The V2 loop of HIV gp120 delivers costimulatory signals to CD4+ T cells through Integrin α4β7 and promotes cellular activation...
... clone UCHL1, BD Pharmingen), anti-Ki67 Alexa Fluor 647 (clone B56, BD Pharmingen), and anti-p24 FITC (clone KC67, Beckman ... Integrin α4β7, is a cell-surface receptor that facilitates CD4+ T cell homing to PPs, MLNs, and LP (7⇓-9). Memory CD4+ T cells ... Human Blood Samples and Primary Cell Preparation.. CD4+ T cells were obtained by negative selection (,95% purity) (Stem Cell ... T cells, we observed a mean of 16% CD25+/Ki67+ cells with anti-CD3 alone vs. 47% CD25+/Ki67+ cells in cultures stimulated with ...
Phosphorylation and negative regulation of CONSTITUTIVELY PHOTOMORPHOGENIC 1 by PINOID in Arabidopsis | PNAS
COP1 S20A, COP1 S21A, and COP1S20 and 21A were cloned into pDONR-221 by BP reaction and then introduced into pEarleyGateway 104 ... D) Firefly LCI assay showing COP1 interacting with PID in tobacco leaf cells. The PID-cLUC and COP1-nLUC constructs were ... COP1 RING-finger, COP1 coiled-coil, and COP1 WD40 domains were amplified from full-length COP1 and cloned into pGEX-4T-1 with ... The target fragments were cloned into the gateway vector pDONR-221 using BP Clonase (Invitrogen). The destination fragments ...