the scans were completed with effective pixel sizes in a range of 12–13 μm by recording 1201 projections over 360 degrees. The reconstruction of the tomograms was set to occur automatically with the dedicated software and performed properly after each measurement. The tumour burden was assessed ...
The s/n-ratio is the ratio between the measurement of the spot intensity and the spot's local background. Gene expression analysis Cells were serum starved for 5 h before treatment with growth factors and inhibitors for the indicated time points. Total RNA was isolated using the High Pure ...
Vascular responses to various vasoactive mediators were applied topically and blood flow was then measured using LSPI. Measurement of vascular maturity was evaluated through immunohistochemical volume analysis of the endothelium and smooth muscle in MCL's using CD31 and alpha-smooth muscle actin antibodies...
MG132 was diluted in DMSO at a stock concentration of 5 mM. For the proteasome activity assay, U937 and SH-SY5Y cells were incubated at indicated concentrations for 2 h before the measurement (see section below). For protein stability assays, U937 cells were incubated for 6 h at 20...
(b) Quantitative measurement of protein–protein interactions using a luciferase-based mammalian two-hybrid assay. HeLa cells stably transfected with a GAL4-driven luciferase reporter and expressing paired GAL4DBD–MCL-1:VP16AD–NOXA, VP16AD–BCL-2:GAL4DBD–BIM or GAL4DBD–BCL-XL:VP16AD–...
This software allows for the measurement of pixel intensities, which can be used to quantify the expression levels of different proteins in IHC-stained samples. To assess the staining intensity, we utilized the IHC Profiler plugin, which is a free download available from the official ImageJ web...
among which the Bcl-2 protein family plays a key role. This family includes anti- and proapoptotic proteins. Cancer cell resistance to apoptosis is commonly associated with overexpression of the antiapoptotic members of Bcl-2 family proteins, in particular, Bcl-2, Bcl-xL, and Mcl-1. Subsequent...
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After a 24-hour incubation, the supernatants were harvested and stored at 70°C until measurement. IL-6 secretion was measured using Duoset ELISA development kits (R&D Systems). All measurements were done in triplicate. Results Seliciclib treatment decreases viability of MM cells in a dose-...
Measurement of ROS levels. Cells were gently harvested, washed with PBS and incubated with 5 μM 2′7′ -dichlorodihydrofluorescein diacetate (H2DCFDA; Molecular Probes) at 37 °C for 30 minutes according to the manufacturer's protocol. Cells were then washed with PBS ...