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A DNA-based molecular probe for optically reporting cellular traction forces.

A DNA-based molecular probe for optically reporting cellular traction forces.

Nature methods (2014-10-13)

Brandon L Blakely, Christoph E Dumelin, Britta Trappmann, Lynn M McGregor, Colin K Choi, Peter C Anthony, Van K Duesterberg, Brendon M Baker, Steven M Block, David R Liu, Christopher S Chen

ABSTRACT

We developed molecular tension probes (TPs) that report traction forces of adherent cells with high spatial resolution, can in principle be linked to virtually any surface, and obviate monitoring deformations of elastic substrates. TPs consist of DNA hairpins conjugated to fluorophore-quencher pairs that unfold and fluoresce when subjected to specific forces. We applied TPs to reveal that cellular traction forces are heterogeneous within focal adhesions and localized at their distal edges.

MATERIALS

Product Number

Brand

Product Description

Sigma-Aldrich

Acetic acid, natural, ≥99.5%, FG

Sigma-Aldrich

Urea-¹²C, 99.9 atom % ¹²C

Sigma-Aldrich

1,1,2,2-Tetrabromoethane, 98%

Sigma-Aldrich

Argon, ≥99.998%

Sigma-Aldrich

Formamide, ACS reagent, ≥99.5%

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Methanol, suitable for HPLC, gradient grade, 99.93%

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Acetic acid, ≥99.5%, FCC, FG

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Propylamine, ≥99%

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Formamide, BioUltra, for molecular biology, ≥99.5% (T)

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Urea solution, BioUltra, ~8 M in H₂O

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Propylamine, purum, ≥99.0% (GC)

Supelco

Methanol, analytical standard

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Propylamine, 98%

Sigma-Aldrich

1,1,2,2-Tetrabromoethane, purum p.a., for separation (of mineral compounds), ≥98.0% (GC)

Sigma-Aldrich

Magnesium chloride solution, BioUltra, for molecular biology, ~0.025 M in H₂O

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Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC

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Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid

Supelco

Ethanol solution, certified reference material, 2000 μg/mL in methanol

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Acetonitrile solution, contains 0.1 % (v/v) formic acid, suitable for HPLC

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Magnesium chloride solution, BioUltra, for molecular biology, 2 M in H₂O

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Methanol, anhydrous, 99.8%

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Formamide, BioReagent, ≥99.5% (GC), for molecular biology

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Hydrocortisone, BioReagent, suitable for cell culture

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L-Thyroxine sodium salt pentahydrate, γ-irradiated, powder, BioXtra, suitable for cell culture

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Magnesium chloride solution, PCR Reagent, 25 mM MgCI₂solution for PCR

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Magnesium chloride solution, for molecular biology, 1.00 M±0.01 M

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Urea solution, 40 % (w/v) in H₂O

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(−)-Riboflavin, from Eremothecium ashbyii, ≥98%

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(−)-Riboflavin, meets USP testing specifications

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(−)-Riboflavin, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥98%