Direkt zum Inhalt
Merck

Cytoskeletal forces during signaling activation in Jurkat T-cells.

Molecular biology of the cell (2014-12-19)
King Lam Hui, Lakshmi Balagopalan, Lawrence E Samelson, Arpita Upadhyaya
ZUSAMMENFASSUNG

T-cells are critical for the adaptive immune response in the body. The binding of the T-cell receptor (TCR) with antigen on the surface of antigen-presenting cells leads to cell spreading and signaling activation. The underlying mechanism of signaling activation is not completely understood. Although cytoskeletal forces have been implicated in this process, the contribution of different cytoskeletal components and their spatial organization are unknown. Here we use traction force microscopy to measure the forces exerted by Jurkat T-cells during TCR activation. Perturbation experiments reveal that these forces are largely due to actin assembly and dynamics, with myosin contractility contributing to the development of force but not its maintenance. We find that Jurkat T-cells are mechanosensitive, with cytoskeletal forces and signaling dynamics both sensitive to the stiffness of the substrate. Our results delineate the cytoskeletal contributions to interfacial forces exerted by T-cells during activation.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Ammoniumpersulfat, ACS reagent, ≥98.0%
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylendiamin, BioReagent, suitable for electrophoresis, ≥99.0%
Sigma-Aldrich
Ammoniumpersulfat, Molecular Biology, suitable for electrophoresis, ≥98%
Sigma-Aldrich
Ammoniumpersulfat, reagent grade, 98%
Sigma-Aldrich
Acrylamid, suitable for electrophoresis, ≥99%
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylendiamin, BioReagent, Molecular Biology, ≥99% (GC)
Sigma-Aldrich
Acrylamid, Molecular Biology, ≥99% (HPLC)
Sigma-Aldrich
N,N′-Methylenbisacrylamid, powder, Molecular Biology, suitable for electrophoresis, ≥99.5%
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylendiamin, ≥99.5%, purified by redistillation
Sigma-Aldrich
N,N′-Methylenbis(acrylamid), 99%
Sigma-Aldrich
Nocodazol, ≥99% (TLC), powder
Sigma-Aldrich
Hydrazin Monohydrat, N2H4 64-65 %, reagent grade, ≥97%
Sigma-Aldrich
Hydrazin Hydrat, reagent grade, N2H4 50-60 %
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylendiamin, ReagentPlus®, 99%
Sigma-Aldrich
Acrylamid, suitable for electrophoresis, ≥99% (HPLC), powder
Sigma-Aldrich
Acrylamid -Lösung, 40%, suitable for electrophoresis, sterile-filtered
Sigma-Aldrich
Hydrazinhydrat -Lösung, puriss. p.a., 24-26% in H2O (RT)
Sigma-Aldrich
Ammoniumpersulfat, BioUltra, ≥98.0% (RT)
Sigma-Aldrich
N,N′-Methylenbisacrylamid -Lösung, suitable for electrophoresis, 2% in H2O
Sigma-Aldrich
Ammoniumpersulfat, BioXtra, ≥98.0%
Supelco
Acrylamid, analytical standard
Sigma-Aldrich
N,N′-Methylenbisacrylamid, suitable for electrophoresis (after filtration or allowing insolubles to settle)
Sigma-Aldrich
Acrylamid, purum, ≥98.0% (GC)
Supelco
Acrylamid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
Acrylamid -Lösung, 40% in H2O
Sigma-Aldrich
Acrylamid, for Northern and Southern blotting, powder blend
Sigma-Aldrich
Ammoniumpersulfat, free-flowing, Redi-Dri, reagent grade, 98%