Human umbilical vein endothelial
cells (HUVEC) and human mesenchymal stem cells (MSC) (Lonza, Walkersville,
MD) were cultured as prescribed by the manufacturer. Chinese hamster
ovary cells harboring Notch “Receiver” and Delta “Sender”
transgenes, [receiver line: CHO-K1-TREx + UAS-H2B-Citrine + CMV-H2B-Cerulean
+ CMV-hNotchECD-Gal4 clone F1; sender line: CHO-K1-TREx + TO-hDll1-mCherry]
both graciously provided by Dr. Michael Elowitz (California Institute
of Technology), were cultured as previously described.34 (link) Human plasma fibronectin (BD Biosciences, Bedford,
MA) was fluorescently labeled using Alexa Fluor 555 NHS ester (Invitrogen,
Carlsbad, CA). Biotinylated fibronectin was obtained from Cytoskeleton,
Inc. (Denver, CO) or made in-house using Biotin-X, SSE, 6-((biotinoyl)amino)hexanoic
acid, sulfosuccinimidyl ester, sodium salt (Sulfo-NHS-LC-Biotin),
(Invitrogen, Carlsbad, CA), and fluorescently labeled using Alexa
Fluor 647 NHS ester (Invitrogen, Carlsbad, CA). Neutravidin and Neutravidin–Oregon
Green 488 conjugate were obtained from Invitrogen. Poly(dimethyl siloxane)
(PDMS; Sylgard 184, Dow Corning, Midland, MI) was used at 10:1 (w:w)
base:curing agent, Young’s modulus ∼1 MPa.
cells (HUVEC) and human mesenchymal stem cells (MSC) (Lonza, Walkersville,
MD) were cultured as prescribed by the manufacturer. Chinese hamster
ovary cells harboring Notch “Receiver” and Delta “Sender”
transgenes, [receiver line: CHO-K1-TREx + UAS-H2B-Citrine + CMV-H2B-Cerulean
+ CMV-hNotchECD-Gal4 clone F1; sender line: CHO-K1-TREx + TO-hDll1-mCherry]
both graciously provided by Dr. Michael Elowitz (California Institute
of Technology), were cultured as previously described.34 (link) Human plasma fibronectin (BD Biosciences, Bedford,
MA) was fluorescently labeled using Alexa Fluor 555 NHS ester (Invitrogen,
Carlsbad, CA). Biotinylated fibronectin was obtained from Cytoskeleton,
Inc. (Denver, CO) or made in-house using Biotin-X, SSE, 6-((biotinoyl)amino)hexanoic
acid, sulfosuccinimidyl ester, sodium salt (Sulfo-NHS-LC-Biotin),
(Invitrogen, Carlsbad, CA), and fluorescently labeled using Alexa
Fluor 647 NHS ester (Invitrogen, Carlsbad, CA). Neutravidin and Neutravidin–Oregon
Green 488 conjugate were obtained from Invitrogen. Poly(dimethyl siloxane)
(PDMS; Sylgard 184, Dow Corning, Midland, MI) was used at 10:1 (w:w)
base:curing agent, Young’s modulus ∼1 MPa.
