Cells were resuspended in SDS lysis buffer (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% (w/v) SDS, 1 mM EDTA, pH 8.0, cOmplete Protease Inhibitor Cocktail (Roche)) and sonicated by a Bioruptor Pico (30 cycles, 30 s on and 30 s off, Diagenode), then centrifuged for 10...
Plants exposed to incidences of excessive temperatures activate heat-stress responses to cope with the physiological challenge and stimulate long-term acclimation1,2. The mechanism that senses cellular temperature for inducing thermotolerance is still unclear3. Here we show that TWA1 is a temperature-s...
at 13.000 r.p.m (pico 17, Thermo). The supernatants were purified for RNA using a high purity viral nucleic acid kit (Roche) according to the manufacturer’s instructions. The purified RNA was analyzed using the RT-qPCR protocols described previously. Statistical analysis The statistical ...
(goat-anti rabbit HRP) at a dilution of 1: 20,000. The blots were developed using SuperSignal West Pico Chemiluminescent Substrate from Pierce, and images were captured on a Kodak image station 4000MM pro from Carestream Health, Inc. To detect actin, the membranes were stripped before being ...
This meta-analysis was performed according to the PRISMA guidelines using the PICO format. We searched in three electronic databases (PubMed, Embase, Cochrane Trials), and assessed heterogeneity using the Q-test and I-squared statistics, meta-regression was also performed. Odds ratios (OR) and ...
Using an example of a well characterized splicing factor, Fox2, we demonstrate that changes in Fox2 expression levels can be used to predict changes in inclusion levels of alternative exons that are flanked by Fox2 binding sites. Conclusions We propose that alternative splicing is an important ...
Units for each entry shown is in picomoles of GTP hydrolyzed per min/pmol of ARL13B ± 1 S.E. of the mean. The concentration of each preparation of ARL13B was 1 μm, and the amount of crude bovine brain ARL13B GAP was also constant at 15 μg of protein, prepared as described ...
Unmyristylated Cα1 was used as reference (data not shown). In the presence of 1 mM ATP and 5 mM MgCl2 the R subunits were simultaneously run over both C subunits on the sensor chip at a flow rate of 30 μL/min. In the case of RIα and RIβ they were run over the sensor ...
Then the sonication of the chromatin was performed with a Bioruptor Pico for four-five rounds of 30 s on and 30 s off cycles. The sonicated chromatin was cleared by centrifugation and the supernatant was used for ChIP. Chromatin was incubated with antibodies pre-bound to Dynal magnetic ...
Specific complexes were detected using the SuperSignal West Pico kit (Thermo Fisher Scientific, Inc.). The following antibodies were used: anti-CILP2 from Santa Cruz, anti-E-Cadherin, anti-N-Cadherin, anti-matrix metalloproteinase (MMP) 9, anti-MMP2 from Abcam, anti-Claudin-1 from Cell ...