Subsequently, these parameters were used for the calculation of the aspect ratio to evaluate the elongation of mitochondria (Aspect ratio = major axis/minor axis). The form factor was defined as (Pm2)/(4πAm), where Pm is the perimeter and Am is the area of the mitochondrion. The ...
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 ...
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...
For the precipitation of proteins from cell culture supernatants, serum-free (OptiMEM) conditioned media of the overnight cultures were used. 10 µL (1/100 of vol.) of 2% sodium deoxycholate (NaDOC) was added to 1 mL sample and was shortly vortexed. After 30 min of incubation on ice,...
Primary human or murine HSCs from wild-type or p70S6K–/– mice as well as LX-2 cells were used for in vitro experiments. Specific small interfering RNA or CEP-1347 were used to silence or inhibit p70S6K and assess its functional relevance in viability, contraction and migration assays, ...
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 ...
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 ...
After washes in TBST, immunoreactivity was visualized using Western blotting detection reagent (SuperSignal West Pico enhancer, Thermo Scientific, Waltham, MA, USA). Expression of GAPDH (monoclonal mouse, Sigma-Aldrich, St. Louis, MO, USA, 1:50,000) was used as loading control. SAMHD1 siRNA ...
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...
Absolute amounts of individual transcripts were calculated and displayed as picograms (pg) of mRNA per 200 ng of total RNA. Each data point represents one biological sample consisting of three flowers. Primers used for the absolute quantification of PhnsLTP1, PhnsLTP2, and PhnsLTP3 ...