correction for bed shear stress

... polymerase (2.5 – U) General PCR program was 94°C for min; 30 cycles of 94°C for 30 s, 60°C for 30 s, 68°C for min/kb (for Pfx, min/kb) of DNA length; 68°C for 10 and PCR was carried out in a thermocycler ... prepared for RNA in situ hybridyzation Primers for amplification of DNA template for in situ hybridyzation are listed in table 2.4 35 Table 2.4 Primers for RNA in situ hybridyzation Gene Forward ... Conditions 36 2.1.4 Transformation of E.coli DH5α Cells 37 2.1.4.1 Preparation of Competent Cells for Heat Shock Transformation 37 2.1.4.2 Heat Shock Transformation of DH5α 37...

Ngày tải lên: 14/09/2015, 08:42

... path for neuronal differentiation for cells exposed to low amounts of BMPs The p300–Smad1 complex is target for yet another regulatory input Neurogenin has been shown to compete with STAT3 for ... decreases both These results stress that BMP signaling is indeed an antiproliferative and differentiation inductive signal for neural stem cells, again (as shown and discussed for murine ES-cells) modulated ... both BMP and TGF-b signaling are dispensable for stemness, even though they are capable of supporting it under specific conditions as described before BMP as an initiator of the germ line Cells...

Ngày tải lên: 19/02/2014, 00:20

... custom-designed microarray platform was employed featuring probes for all Drosophila genes for which different mRNA isoforms generated by alternative splicing have been described [51] Three biological ... by representsfor removal the of enriched changes inreguways.biological of (top) for alternative and wingless insulinfor latedof promoters andlevel bywingless (bottom) by genes that for eachtablevalues ... changes in particular isoforms Quantitative RT-PCR assays were carried out for 15 different genes, of which 11 (70%) were validated Figure shows the results obtained for six of these genes and...

Ngày tải lên: 14/08/2014, 21:20

... and cycling parameters for glyceraldehyde3-phosphate dehydrogenase (GAPDH) were as described previously [17] Primer pairs for ICAM-1 (Accession No: BC015969) (5¢fi3¢) were (forward) CCG TGT ACT ... responsible for the recruitment of inflammatory cells from the blood to the airways, but also acts as a receptor for many viruses that may exacerbate asthma [16] Numerous studies suggest a role for NF-jB ... receptor for infection by rhinoviral and respiratory syncytial viruses, which both increase exacerbations in asthma [28,29] The regulation of ICAM-1 production is therefore a potential target for...

Ngày tải lên: 07/03/2014, 15:20

... cytotoxic stresses, including oxidative stress and UV irradiation, and play an essential role in stress- induced apoptosis [40,41] Therefore, it is likely that Hsp105a and Hsp105b suppress oxidative stress- induced ... have been implicated as key regulators in some forms of stress- induced apoptosis [30,31] Phosphorylation of JNK at Thr183 and Typ185 is required for JNK activation [32] Activation of p38 MAPK ... such an extent that the reaction becomes unproductive for folding [60] Therefore, Hsp105a and Hsp105b seem not to simply suppress oxidative stress- induced apoptosis by regulation of Hsp70-mediated...

Ngày tải lên: 16/03/2014, 06:20

... been used for various purposes for a long time For example, they have been used as reagents for Western blotting, immunostaining, immunoprecipitation and blocking of protein function Therefore, if ... products for VH region were classified into two forms containing the native VH leader signal sequence at the 5¢ end of the VH fragment (SVH form) or no VH leader signal sequence (VH form) The ... stimulation was performed in the presence of exogenous IL-4 In each experiment, cells were cultured for 48 h, then 10 lM BrdU was added to the T and B-cell cultures The cells were reincubated for an additional...

Ngày tải lên: 16/03/2014, 14:20

... 24 h, cells were treated with dimethylsulfoxide (vehicle control), forskolin, cholera toxin (CTX) or PMA for 24 h, before fixing for fluorescence microscopy Scale bar = 25 lm F GFP-LRH15KR Fig Multiple ... cells were fixed in 4% paraformaldehyde for 10 and then mounted For reagent treatments, 24 h after the transfection, media were replaced with fresh media and then 100 lm forskolin (Sigma) in dimethylsulfoxide, ... 13-acetate (Sigma) in dimethylsulfoxide were added Cells were cultured for 24 h before being fixed in 4% paraformaldehyde for 10 and mounted Images were obtained with a Zeiss Axioskop2 microscope...

Ngày tải lên: 23/03/2014, 06:20

... performed as previously described [22] Briefly, total RNA was reverse-transcribed using 500 ng poly-dT and 0.25 U of avian myeloblastosis virusreverse transcriptase at 42°C for 75 and 95°C for ... and LH for h; FSH(+): COCs were cultured with FSH and LH for h; EGF(-): COCs were cultured without EGF for h; EGF(+): COCs were cultured with EGF for h; +H89: COCs were cultured with H89 for h; ... FSH and LH for 2.5 h; Cont: COCs were cultured with FSH and LH for 2.5 h; H89: COCs were cultured with FSH, LH and H89 for 2.5 h; SB: COCs were cultured with FSH, LH and SB203580 for 2.5 h; LY:...

Ngày tải lên: 20/06/2014, 07:20

... to Dct was performed as described [9,15] TUNEL assays were performed following the manufacturer’s guidelines (Roche Diagnostics, Basel, Switzerland) Standard protocols were used for tissue processing ... expression during eye formation; and Additional data file 2, a figure showing the expression of TGF␤ isoforms during eye formation Acknowledgements We thank B Langsam and C Imsand for their excellent ... Tgfbr2-mutant mice The generation of mice used in this study has been described before [9,11,16,41] Briefly, loxP-sites for Cremediated recombination were introduced into the mouse Tgfbr2 locus...

Ngày tải lên: 06/08/2014, 18:21

... might result in increased receptor avidity for otherwise low-affinity foreign or even unrelated self TLR ligands Another Page of 11 (page number not for citation purposes) possibility is that ... receptor for antigen (BCR)-mediated signaling For example, blocking the calcineurin pathway with cyclosporine A diminished BCR-dependent proliferation, whereas treatment Page of 11 (page number not for ... structural evidence for antigen selection of anti-DNA antibodies Annu Rev Immunol 1994, 12:487-520 34 Tan EM: Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology...

Ngày tải lên: 09/08/2014, 07:20

... Shen Han-ming for their suggestions and advice My heartfelt thanks to Dr Lim Shen Kiat and Dr Shirly Chong for their ready help and invaluable advice whenever needed and Ms Lee Huiyin for all the ... Lim Shen Kiat and Emma May Stanford for taking their precious time to proof-read this thesis I would like to thank all the past and present members of YPL lab for the wonderful working experience, ... sincere thanks to my family, especially my parents for their constant moral support, my sister for her listening ears and my little brother for his IT expertise and help in drawing the diagrams...

Ngày tải lên: 11/09/2015, 10:00

... differentiation For the ErbB family, our target was to investigate the interactions between the receptors before ligand binding, which are crucial for the activation of the receptors For the Cdc42 ... fluorescent labels, i.e two labels must be close enough to form fluorophores (for complementary methods) or to transfer energy from donor to acceptor (for FRET distance

Ngày tải lên: 14/09/2015, 14:25

... inhibitory eNOS-caveolin complex reforms 51 However, the activation of eNOS can also be regulated by phosphorylation For example, an increase in blood flow, and thus in shear stress, as well as hypoxia ... present in endothelial cells is activated by fluid shear stress in a tyrosine kinase-dependent manner 74 It is also possible that the shear force may induce the endothelium to release some active ... pathways for DAG formation PLD, phospholipase D; PLA2, phospholipase A2 G-3-P, glyceraldehydes-3-phosphate The de novo synthesis pathway is mainly responsible for hyperglycemia-induced DAG formation...

Ngày tải lên: 15/09/2015, 17:09

... region For pcDNA-Ng construction, pcDNA3.1 plus (Invitrogen, USA) vector was used for construction The vector map for pEGFP-C2 and pcDNA3.1 plus were shown in Fig.5 and Fig.6 The process for construction ... - 36 1.2 Gel extraction - 41 1.3 Ligation and transformation - 41 1.4 Competent cell preparation for transformation - 41 1.5 Screening of positive clones ... al, 1999) The oxidized Ng forms intramolecular disulfide bonds as detected by increased migration on SDS-PAGE Among the cysteines, Cys 51 is critical for - 13 - disulfide formation and the relative...

Ngày tải lên: 05/10/2015, 22:32

... region For pcDNA-Ng construction, pcDNA3.1 plus (Invitrogen, USA) vector was used for construction The vector map for pEGFP-C2 and pcDNA3.1 plus were shown in Fig.5 and Fig.6 The process for construction ... - 36 1.2 Gel extraction - 41 1.3 Ligation and transformation - 41 1.4 Competent cell preparation for transformation - 41 1.5 Screening of positive clones ... al, 1999) The oxidized Ng forms intramolecular disulfide bonds as detected by increased migration on SDS-PAGE Among the cysteines, Cys 51 is critical for - 13 - disulfide formation and the relative...

Ngày tải lên: 06/10/2015, 20:35

... double-distilled water and stored at -40℃ for use Immediately before use, the solution was centri- http://www.medsci.org Int J Med Sci 2011, 58 fuged at 18000 g for 10 and the supernatant was filtered ... of 50 μl of mixture used for amplification The products were sequenced by Shanghai Sangong Co., Ltd immediately after purification The primers for sequencing were those for amplification Using ... overnight On the next day, the supernatant was obtained and vigorously oscillated for 10 followed by centrifugation at 12000 g for 10 The supernatant was collected and the sediment was re-suspended in...

Ngày tải lên: 25/10/2012, 11:18

... conditions used for the PCR were as follows: 94°C for min, 30 cycles of 94°C for 45 sec, 68°C for 45 sec, and 74°C for min, with final extension at 74°C for The integrity of the RNA used for RT-PCR ... quantification cycling protocol was used: 50°C for minutes, followed by 95°C for 15 minutes, and 45 cycles of 76°C for 30 seconds, 94°C for 15 seconds, and 56°C for 35 seconds JNK, PTEN, and caspase-3 ... 95°C for minutes to inactivate Quantiscript reverse transcriptase An aliquot of each finished reverse transcription reaction was added to the real-time PCR mix A probe for GAPDH was used for normalization...

Ngày tải lên: 31/10/2012, 15:12

... µg/ml pepstatin A; and sonicated for 15 sec Cell debris was removed by centrifugation at 1400 x g for 30 at °C Protein concentration was determined by the Bradford assay [23] Protein kinase phosphorylation ... the basis for further studies addressing the pathophysiological mechanisms of exposure to low concentration of ethanol in HepG2 cells, and provides information about potential targets for the treatment ... Hampong M, et al Stress- induced inhibition of ERK1 and ERK2 by direct interaction with p38 MAP kinase J Biol Chem 2001;276:6905-8 23 Bradford MM A rapid and sensitive method for the quantitation...

Ngày tải lên: 31/10/2012, 15:37

... re-amplified and re-sequenced for confirmation Surveyor nuclease digestion Exon (transcript variant 3) and exon 10 were amplified from selected samples with PCR parameters as described above After a control ... samples, denatured at 95°C for minutes and subsequently re-hybridized In case of sequence deviations in the wildtype samples from the reference sequence this resulted in the formation of heteroduplexes ... has no consequence for the hCDC4 protein Sequence analysis of hCDC4 exon 10 identifies a new intronic SNP During the analysis of exon 10 and adjacent intronic sequences we could for the first time...

Ngày tải lên: 31/10/2012, 16:49

... 10μM for the indicated times The PKA activator, forskolin (Sigma) was used at a concentration of μM for the indicated times Immunofluorescenct confocal microscopy As previously described (4), for ... analysis for claudin-1 as described above Invasion Assays Matrigel invasion assays were performed using transwell migration chambers (Corning life Sciences, Lowell, MA) as previously described (21) ... probed with antibodies and then visualized using the ECL system (Amersham Biosceinces, Piscataway, NJ) For immunoprecipitation, 250μg of lysate was incubated with protein A/G agarose (Sigma) for...

Ngày tải lên: 03/11/2012, 11:17