sleeping beauty mediated transposition in stem cells pdf

... Saikosaponins induce intracellular ROS accumulation in Siha cells, A549 cells, and SKOV3 cells Siha cells, A549 cells, and SKOV3 cells were treated with saikosaponin-a (10 μM) or saikosaponin-d (2 ... described in (A) induced by saikosaponin and cisplatin co-treatment, we first detect morphological changes in saikosaponin and cisplatin-cotreated HeLa cells by acridine orange/ethidium bromide staining ... Saikosaponins induce intracellular ROS accumulation in HeLa cells HeLa cells were treated with cisplatin (8 μM) or saikosaponin-a (10 μM) or saikosaponin-d (2 μM) individually or combination of...

Ngày tải lên: 10/08/2014, 10:20

... variety of cell types, including hepatocytes, to long-chain saturated fatty acids (SFAs) led to increased expression of proinflammatory cytokines, inhibition of insulin signaling, induction of endoplasmic ... Pharmigen, San Diego, CA, USA) binding and PI staining were performed according to [44] Following incubation with 1.0 lgÆmL)1 annexin V and 4.0 lgÆmL)1 PI for 20 min, cells were analyzed by flow cytometry ... results that TAG synthesis was initially hindered following stearate administration and was completely interrupted at longer incubation periods Interestingly, the interruption of TAG synthesis...

Ngày tải lên: 14/02/2014, 22:20

... SB-431542 to block activin ⁄ TGF-b signaling in murine stem cell cultures Both experiments resulted in a strong growth inhibition of the stem cells but, fascinatingly, did not interfere with their ... maintain self-renewal of human embryonic stem cells Stem Cells 22, 770–778 Ying QL, Nichols J, Chambers I & Smith A (2003) BMP induction of Id proteins suppresses differentiation and sustains ... BMP signaling is indeed an antiproliferative and differentiation inductive signal for neural stem cells, again (as shown and discussed for murine ES -cells) modulated by LIF ⁄ STAT3 in a highly...

Ngày tải lên: 19/02/2014, 00:20

... fused to d1-EGFP are maintained in the cells by binding to antigen Generation of soluble binding-defective and cysteine-deleted mutants To demonstrate that the observed intracellular colocalization ... was obtained with scFv 4A-d1-EGFP and 7A-d1-EGFP expressed in parallel Interestingly, in the case of 4A (but not 7A), we found a minor percentage of cells with a faint cytoplasmic staining (arrow) ... the 4A- or the 7A-cotransfected cells, thus demonstrating that strong in vivo binding of the destabilized 13R4-d1-EGFP proteins allows them to be maintained in the cells To rule out the possibility...

Ngày tải lên: 19/02/2014, 18:20

... grown in Dulbecco’s modified Eagle’s medium containing 5% fetal bovine serum and 1% nonessential amino acids and suspended in 50 mM Tris/HCl pH 7.5, containing 10 lgÆmL)1 aprotinin The cells were ... angiotensin IV and may play a role in the maintenance of memory [19] In the current study, we detected the P-LAP protein in the kidney and examined its potential role We have observed that 8-arginine-vasopressin ... aprotinin) After determination of the protein concentration, cell lysate containing 100 lg of protein was diluted to 300 lL with lysis buffer, added 15 lL of immobilized streptavidin (6% cross-linked...

Ngày tải lên: 08/03/2014, 02:20

... inactive [49] In our experiments NDGA was effective in inhibiting the activity of purified recombinant MTE-I in an in vitro assay performed in a Hepes-based buffer containing only KCl Information ... known that an acyl-CoA binding protein (ACBP) known also as DBI (diazepam binding inhibitor) is expressed in high concentrations in specialized cells such as steroid producing cells of the adrenal ... performed in triplicate activity in vitro Figure shows that a 5-min preincubation of recombinant protein with NDGA inhibited thioesterase activity Interestingly, NDGA produced only a 20% inhibition...

Ngày tải lên: 08/03/2014, 09:20

... ERK1 ⁄ and c-Jun N-terminal kinase pathways, through AP-1 signaling [26] AP-1 family proteins are good candidates for mediating NGF-induced miR-221 ⁄ 222 expression in PC12 cells A previous study ... new insights into the MAPK signaling pathway 3274 Experimental procedures Cell culture and transfection PC12 cells were maintained in Dulbecco’s modified Eagle’s medium plus 10% fetal bovine serum, ... had increased by approximately 26-fold NGF induced sustained activation of ERK1 ⁄ over this time course (Fig 1D) We further examined whether EGF stimulation also induces miR-221 ⁄ 222 in PC12 cells...

Ngày tải lên: 16/03/2014, 01:20

... MA-10 cells were labeled for h at 37 °C with [1-14C] AA (1 lCiÆmL)1 per · 106 cells) in serum-free media containing 0.5% fatty acid-free bovine serum albumin Then, cells were incubated in either ... antisense transfected cells lation of steroidogenesis using 300 lm AA in the presence of albumin These different results can be explained by the fact that protein binding can increase the overall ... present in an aqueous environment by effectively decreasing the insoluble fraction [2] Albumin, in particular, binds specifically to fatty acids [33] In our experiments, we added albumin in the...

Ngày tải lên: 16/03/2014, 12:20

... that: (a) DIFs increase [Ca2+]c in some tumor cells [9–12], (b) DIFs activate Akt ⁄ protein kinase B in human leukemia K562 cells [13], (c) DIF-1 inactivates STAT3 in gastric cancer cells [15], ... consumption in a way other than by increasing [Ca2+]c or inhibiting PDE1 To support our hypothesis that an increase in [Ca2+]c is not involved in the DIF-1-promoted glucose consumption, we examined ... supplemented with 75 lgÆmL)1 penicillin, 50 lgÆmL)1 streptomycin and 10% (v ⁄ v) fetal bovine serum RGM-1 cells were maintained in vitro at 37 °C (5% CO2) in DMEM containing a low concentration (1000...

Ngày tải lên: 23/03/2014, 09:20

... nonactivating fragment (154–279, CIP) that inhibits Cdk5 activity and binds with high affinity (C) Inhibition of Cdk5 activity by CIP Cdk5 kinase activity was determined by preincubating various ... markedly inhibited in vitro by less than lM CIP CIP inhibits the activity of Cdk5 kinase in transfected HEK293 cells To explore the inhibitory effects of CIP on the Cdk5/p25 complex activity in vivo, ... (Fig 2B, c and d, respectively) in the transfected cells To determine whether CIP inhibits the protein phosphorylation activity of Cdk5/p25 in transfected cells, kinase activity assays were performed...

Ngày tải lên: 23/03/2014, 21:21

... sections were incubated for 30 minutes at room temperature in streptavidin–peroxidase complex (1:500 in DPBS; Dako), rinsed in distilled water and incubated in fast-DAB (3,3'-diaminobenzidine) peroxidase ... Khan and T.E Hardingham, unpublished data) Mammalian cells are normally cultured in air (containing 20% oxygen) with 5% carbon dioxide added, but some cells, including adult stem cells, have been ... medium contained basic medium (as above, but without serum) with × insulin–transferrin– selenium supplement (ITS+1; final concentrations 10 μg/ml bovine insulin, 5.5 μg/ml transferrin, ng/ml sodium...

Ngày tải lên: 09/08/2014, 10:20

... healing in animal models [13-15] Recombinant human BMP-2 and BMP-7 are used clinically in spinal fusion and the healing of tibial fractures To obtain a clinically acceptable result, these proteins ... application of chordin also limits in vitro osteogenesis [19] Overall, these findings confirm that BMP-2 is important in driving and sustaining osteogenic differentiation in this in vitro model This ... BMP-2 and chordinstem cells during osteogenic differentiation of human mesenchymal stem cells (a) Temporal progression of bone morphogenetic protein (BMP)-2 and chordin expression during osteogenic...

Ngày tải lên: 09/08/2014, 10:23

... aminoethylcarbazole staining (red) for active caspase-3 (hematoxylin-counterstained) before (a) and after (b) treatment and diaminobenzidine staining (brown) for TUNEL (hematoxylin-counterstained) ... significant Results Clinical response following intra-articular glucocorticoids is accompanied by a decrease in the number of ST T cells All patients included in the study were clinical responders ... fluorescent staining of CD3+ cells (red, rhodamin red) before (a) and after therapy (b), TUNEL+ cells (green, fluorescein) before (c) and after therapy (d) (b, e), and superimposed stainings before...

Ngày tải lên: 09/08/2014, 13:22

... lesser extent) NC2 binding, thereby providing a model for binding of GTFs in the absence of core elements and alterations in TFIIB/NC2 ratios inside cells In addition to defining a library of promoters ... and TFIIB binding in vivo The high prevalence for the occurrence of motif combinations in TFIIB-dominated genes in illustrated in Figure 5e In line with the known synergy between INR and TATA ... calls is indicated by the dotted horizontal line The red dots indicate the average expression in gene groups with increasing TFIIB occupancy They were determined by moving a sliding window (step...

Ngày tải lên: 09/08/2014, 20:21

... L-g-glutamyl-L-cysteinyl-glycine, a ubiquitous endogenous antioxidant It plays an important role in maintaining intracellular redox equilibrium and in augmenting cellular defenses in oxidative stress [20,21] In ... was observed in HepG2 cells It is then very clear that HIF-1a proteins in hypoxic cells were significantly decreased with BSO concentrations gradually increasing In addition, the inhibition of ... glutathione is in the reduced form Shifting redox equilibrium is in favor of a reducing or oxidizing state; that is in modification of the redox status in cells [22,23] The g-glutamylcysteine sythetase...

Ngày tải lên: 10/08/2014, 10:21

... through the inhibition of IκB kinase Page of 15 [13,18] There is increasing interest in curcumin as a therapeutic option for OA and RA, with evidence that curcumin inhibits the IL-1β-induced activation ... containing viable, rounded cells that were embedded in a cartilage specific matrix In these Page of 15 cultures, a four-hour curcumin pre-treatment (d) was as effective in inhibiting IL-1β induced ... signalling molecules such as β1-Integrin Similar observations have been made in chondrocytes [15] Interestingly, integrins have already been shown in several tissues to be able to mediate curcumin...

Ngày tải lên: 12/08/2014, 14:22

... Medicine 2009, 7:6 http://www.translational-medicine.com/content/7/1/6 CD34+ Figure stem cells transdifferentiated in muscle cells in vitro CD34+ stem cells transdifferentiated in muscle cells in ... amplified in co-cultures, indicating the absence or very low expression of dystrophin in co-cultures (data not shown) Transdifferentiation of CD34+ stem cells into muscle cells During the expansion ... suggests that the fusion of stem cells and muscle cells was sufficient to induce the stem cells nuclei to express muscle cells proteins, restoring the absent dystrophin expression More than different...

Ngày tải lên: 18/06/2014, 15:20

... differentiate into teratocarcinomas, three germ layer tumours containing a small proportion of undifferentiated stem cells [13] In contrast, nullipotent 2102Ep EC cells can avoid differentiation during ... towards increased expression of mature miRNAs in 2102Ep cells The miRNAs and levels of expression are listed in Additional file altered in 2102Ep cells following RA treatment but not in NTera2 cells ... in undifferentiated 2102Ep cells but not in NTera2 cells remain unaltered upon RA treatment of 2102Ep cells These 21 miRNAs represent an independent miRNA mechanism employed by 2102Ep cells in...

Ngày tải lên: 20/06/2014, 07:20