To assess the effect of B:C ratio on T cell expansion, PBMCs were seeded at a concentration of 250 × 103 cells/mL in a 24-well culture plate (based on the results of seeding density comparative groups), and were then stimulated once with microbeads at different B:C ratios (1:10...
At this point, we used a 4-min incubation with Accutase (Sigma, cat. A6964) to detach hiPS cells and subsequently counted and seeded cells onto Geltrex (Thermo Fisher, cat. A1413301)-coated 12-well plates at a density in the range of 3–7 × 105 cells per well in iPS-Brew + 2...
Mixture ofα-syn monomers and fibrils exacerbates cell death in hippocampal primary neurons. (a–f) Hippocampal primary neurons were plated in 96-well plate and treated with Tris buffer orα-syn species for the indicated times. (a–e) Cells were stained with Sytox Green (SG), a membrane-im...
Another explanation may be related to the physiologically higher innervation density in proximal vs distal skin29, with consequently higher amount of tau and higher propensity to seed/aggregate. This might also explain the lower aggregation activity in PD in cervical compared to MSA and HC; indeed,...
4a). These fractions with GGT-tau of different aggregation states were subsequently tested for their seeding competency in the tau biosensor cell line. Interestingly, the FRET tau seeding assay detected differences in FRET density across different tau species isolated from the same sample. P3 GGT-...
This overnight culture was expanded 1:100 into terrific broth medium and grown to an optical density (OD) of 0.6–0.8 at 37 °C with shaking. Protein expression was induced by the addition of 0.5 mM isopropyl β-D-1-thiogalactopyranoside at room temperature for 16 h. Cells were ...
For whole cell lysate, bacteria grown on YESCA agar at 26 °C for 48 h were suspended in 50 mm potassium Pi, pH 7.2, normalized by optical density at 600 nm (A600), and pelleted down. The pellets were pre-treated with or without 70 μl of HFIP immediately followed by SpeedVac ...
Transduction of biosensor cell lines, flow cytometry and seeding analyses The seeding assay was conducted as previously described [14] with the following changes: v2H cells were plated 20 h before seed transduction at a density of 16,000 cells/well in a 96-well plate in a media ...
Although RT-QuIC demonstrates unparalleled detection sensitivity and specificity for brain and lymphoid tissues, it has lower sensitivity for other sample types; this has in-part been ascribed to lower prion density and RT-QuIC reaction inhibitors16. Importantly, the inhibitory effect of certain ...
Richter transformation (RT) is a paradigmatic evolution of chronic lymphocytic leukemia (CLL) into a very aggressive large B cell lymphoma conferring a dismal prognosis. The mechanisms driving RT remain largely unknown. We characterized the whole genome,