yield choice of culture dish and cell seeding density

... cell death Type III or necrotic cell death is characterized by rapid loss of plasma membrane integrity, cellular swelling and subsequent release of internal contents [48] In C elegans, gain -of- function ... Autophagy and signaling: their role in cell survival and cell death Cell Death Differ 12(Suppl 2), 1509–1518 Meijer AJ & Codogno P (2006) Signalling and autophagy regulation in health, aging and disease ... threonine 119 and promotes the dissociation of beclin from Bcl-XL and the induction of autophagy, similar to the mechanism by which JNK1 regulates autophagy by phosphorylation and dissociation of beclin...

Ngày tải lên: 29/03/2014, 08:20

... accumulation of amino-acid pools, mitochondrial function, and the insulin and TOR pathways in C. elegans longevity Black lines indicate the activating or repressive function of members of different ... pathway The physiological role of TOR kinase is to sense nutrient levels - such as cellular amino-acid pools - and to regulate transcription and protein biogenesis and degradation accordingly TOR ... downregulation of TOR increases longevity; ife-2 is an effector of the TOR pathway and animals carrying a mutation in this gene also live longer; and ife-2 mutants accumulate pools of amino acids...

Ngày tải lên: 06/08/2014, 19:21

... loss of function mutant strains of lin-4 and let-7 For example, progeny of animals injected with antisense oligonucleotides against lin-4 show delayed exit in differentiation of seam cells and ... solubility in water and shows increased cellular uptake and availability The authors also modified these oligonucleotides by conjugating one to three molecules of rhodamine per molecule of dextran These ... inhibition of small RNA function PLoS Biol 2004, 2:e98 doi:10.1186/jbiol230 Cite this article as: Kundu ST and Slack FJ: Robust and specific inhibition of microRNAs in Caenorhabditis elegans Journal of...

Ngày tải lên: 06/08/2014, 19:21

... mixtures were spotted onto PEI cellulose sheets with lL of nonradioactive tracer (100 lg of 5-FU and 100 lg of uridine for the UP assay mixture; 100 lg of 5-FU and 100 lg of deoxyuridine for the TP ... tests of wild-type and upp-1(jg1) worms Fig S2 Lifespan tests of wild-type worms and three upp-1 mutants Fig S3 Sequence alignments of C elegans UPP-1 and human UPP-1 Fig S4 Sequence alignments of ... reversible phosphorolysis of uridine, yielding uracil and Rib1P, and is an important enzyme in the pyrimidine salvage pathway Human UP and TP each exhibit both uridine and thymidine phosphorylase...

Ngày tải lên: 30/03/2014, 01:20

... activation of MIG-17 ⁄ ADAMTS S Ihara and K Nishiwaki staining of cross-sections of L3 larvae with antibody to GFP revealed signals in the cytoplasm of muscle cells, the source of MIG-17 expression, and ... detected a faint band of the MIG-17–6His proform and an additional lower molecular weight band in worm lysates (Fig 3B) Monoclonal specifically recognized the activated forms of MIG-17–GFP and MIG-17–6His ... proform localizes to the surfaces of the gonad, intestine and hypodermis corresponding to the basement membranes [17] To determine the sites of activation of MIG-17 and tissue distribution of...

Ngày tải lên: 30/03/2014, 04:20

... standard Z-score (l - m)/s where l is the observed number of links in the subnetwork, and m and s are the mean and standard deviation of the number of links across 1,000 random collections of ... Jeff Wrana, and Brenda Andrews for helpful comments on the work and the manuscript We thank Jason Moffat for efforts at the proof -of- principle stage and thank Theresa Stiernagle and the C elegans ... levels of agreement The significance of the LOFA score was assessed by generating million random LOFA values We first constructed a random dataset in which the genes associated with loss -of- function...

Ngày tải lên: 06/08/2014, 18:21

... embryonic cell cultures allow the collection of cells at early stages of development, methods for the separation, culture and collection of fully developed tissues have not been established and might ... patterns of subsets of neurons, in combination with analyses of neural functions, might provide insights into understanding the distinct roles of cells within the neural network reports Materials and ... instead of FITC Forty-six wild type and 56 che-2::PABP worms at L4 to young adult were observed Cells were identified by their positions and the percentage of stained cells was scored Responses of...

Ngày tải lên: 14/08/2014, 14:21

... this time period in the context of the rate of change of coding genes, C elegans and C briggsae only share about 60% of their genes as 1:1 orthologs, and a full 10% of genes encoded in either genome ... evidence for the redundant functions of duplicated genes comes from the analysis of lossof-function phenotypes of single genes; in both yeast and worms, inactivation of a duplicated gene is less likely ... into each well of a 96-well flat-bottom plate and 40 µl of the resuspended bacterial cultures were added For combinatorial RNAi feeding experiments, resuspended saturated cultures of different...

Ngày tải lên: 14/08/2014, 17:22

... subcellular organelle that projects from the surface of many eukaryotic cells in vertebrates, including kidney and endothelial cells, myocardial cells, odontoblasts, retinal photoreceptor cells ... level of expression in the ciliated subset of neuronal cells versus predominantly non-ciliated cell subsets (that is, pan-neuronal, intestinal and muscle cell subsets) Combining the X-box and SAGE ... phasmid Cell body Dendrite Dendrite Cell body Dendrite Cell body F32A6.2 - amphid Cell body F32A6.2 - phasmid Dendrite Cell body F41E7.9 - phasmid Dendrite Cell body Dendrite F41E7.9 - amphid Cell...

Ngày tải lên: 14/08/2014, 17:22

... spontaneous release of ATP, probably because of influx of cADPR through CD38 and subsequent activation of intracellular ryanodine-sensitive cADPR receptors The EFS-evoked release of ATP, however, ... the peaks of b-NAD+ + ADPR + cADPR, AMP and Ado represented the amounts of endogenously formed nucleotides and nucleosides plus products of the degradation of the exogenous cADPR, and therefore ... both CD38+ ⁄ + and CD38) ⁄ ) mice was insensitive to inhibition of fast Na+ channels with TTX, inhibition of connexin and pannexin hemichannels with CBX and FFA, and cleavage of SNAP25 with BoNTA...

Ngày tải lên: 05/03/2014, 23:20

... helix of the Dyb-1 coiled-coil domain strongly decrease or abolish the functional properties of Dyb-1 DISCUSSION The aim of this study was twofold: first, to confirm and refine the localization of ... plasmids and AD-Dyb-1 plasmids (AN450, AD-2¢5, AD-5¢1, AD-5¢2B, AD-5¢5B, or AD-6¢4) Overnight cultures (5-mL) were prepared in SD media minus tryptophan and leucin The next day, mL of overnight culture ... 2.9 kb of upstream sequence, and various restriction enzyme sites were removed and added by performing synonymous point mutations to yield the construct dyb-1:gfp VII, which has single AgeI and...

Ngày tải lên: 08/03/2014, 23:20

... where A and B are the MICs of drug A and drug B in the combination, MICA and MICB are the MICs of drug A and drug B alone, FICA and FICB are the FICs of drug A and drug B and n is the number of wells ... existence of a direct correlation between the peptide dose and the extent of both microbial death and membrane disturbance 10 Fig Effect of Esc(1–18) on the permeation of the E coli ATCC 25922 IM Cells ... membrane was noted after the first of peptide treatment, and this was followed by more damage and loss of cellular integrity, with a partial discharge of the cellular contents, within 20 (Fig...

Ngày tải lên: 16/03/2014, 00:20

... Localization of GLUT8 is not altered in cells lacking AP3 subunits (mocha and pearl cells) In order to confirm our biochemical data, we investigated the subcellular localization of GLUT8 in living cells ... proteins LAMP1 and LAMP2 in spermatocytes and spermatids The EXXXLL motif interacts with AP1 and AP2 but not with AP3 or AP4, and appropriate targeting of GLUT8 is dependent on both AP1 and AP2, while ... discrepancies in order to (a) identify the subcellular localization of GLUT8 in testis, (b) elucidate the role of APs in GLUT8 sorting, and (c) understand the role of the EXXXLL motif in GLUT8 sorting...

Ngày tải lên: 16/03/2014, 02:20

... indicated times Cells in the absence of any of these were taken as the control (Con) Two micrograms of total RNA were reverse transcribed with random primers and the levels of PEPCK and G6Pase mRNA ... Each band was analysed densitometrically and the values are depicted after normalization of PEPCK and G6Pase bands with those of 18S rRNA (B) Each point is the mean ± SEM of three sets of experiments ... treatment (50 nM, 15 min) Cells incubated in the absence of any of these were taken as the control (Con) On termination of incubation, cells were lysed and the nuclear (50 lg) and cytosolic (40 lg)...

Ngày tải lên: 16/03/2014, 02:20

... number of offspring (250–300 progeny); (d) growth on a solid surface medium; and (e) transparent body allowing the observation of cells in mature and developing animals Furthermore, as about 60% of ... A' E B F C G D E' H Fig Analysis of expression pattern of gstk-1 and gstk-2 in embryo and adult C elegans Projection of confocal images of GFP::GSTK-1 (A–D) and GSTK-2::GFP (E–H) at different ... that, in human cells, hGSTK1 is both peroxisomal and mitochondrial and contains a C-terminal PTS1 [4] This preserved intracellular localization of GST kappa of both nematodes and vertebrates,...

Ngày tải lên: 16/03/2014, 03:20

... and in adult worms (Fig 1A) At higher magnification, we saw GFP staining within pharynx cells of the procorpus, metacorpus, isthmus and terminal bulb, and extracellular staining in the lumen of ... ALIGN_000543) For identification and annotation of protein domains and the analysis of domain architectures the tools of the SMART [33], PFAM [26], ProDom [34] and INTERPRO [35] protein domain ... only in the head of adult hermaphrodites, but not within pharynx cells (Fig 1C) The expressing cells are located outside of the pharynx, around the metacarpus and the terminal bulb, and could include...

Ngày tải lên: 23/03/2014, 15:21

... 500 spectrometer and was of the order of 15 h The 13C–1H coupling constant was 140 Hz, with a sweep width in the F2 (1H) dimension of 10.0 p.p.m and in the F1 (13C) dimension of 230 p.p.m Water ... resonance of 3.56 p.p.m and a 13C resonance of 42.4 p.p.m., in excellent agreement with the glycine standard A 13C-1H HMBC experiment (Fig 3B) produced a cross-peak at 1H resonance at 3.56 p.p.m and ... p.p.m consistent with the presence of a substituent at the 7-position of HepII Signals indicative of the presence and absence of substituents at the 7-position of HepII in the L4 galE oligosaccharide,...

Ngày tải lên: 23/03/2014, 21:21

... youthful and active longer than normal, suggesting that loss of the germline precursor cells extends lifespan because it slows the animals’ rate of aging Thus, in these animals, the germ cells not ... reproductive system contains only four cells, so using a laser beam to ablate either the precursors of the germline (Z2 and Z3) or the precursors of all of the reproductive tissues (Z1–Z4) with ... can be devoted to cell and tissue maintenance This idea, that there is a “cost of reproduction,” has been put forth by evolutionary biologists, and there are many examples of reciprocal relationships...

Ngày tải lên: 28/03/2014, 16:20

... composed of three major structures, a central cell body and two networks of extensively branched membrane structures, the dendrite and the axon Growing axons respond to a wide variety of extracellular ... (a) Genetic and physical maps of chromosome III in the vicinity of the unc-69 locus unc-69 is close to and left of ced-9 Cosmids and subclones able to rescue the locomotion defect of unc-69(e587) ... and pRH143 to create LexA-bait and GAD-prey plasmids pRE192 and pRH143 are derivatives of pBTM116 and pGAD424, respectively, and are gifts of K Basler (University of Zurich) Single-amino-acid changes...

Ngày tải lên: 06/08/2014, 18:21