disposable ecis arrays  (Applied BioPhysics)

Journal: Molecular Vision

Article Title: Elucidating the molecular basis of PPCD: Effects of decreased ZEB1 expression on corneal endothelial cell function

doi:

Figure Lengend Snippet: The impact of ZEB1 reduction on cell migration in HCEnC-21T cells. A : HCEnC-21T cells were transfected with either ZEB1 siRNA-C or scrambled siRNA (sc-siRNA), and knockdown of ZEB1 was validated with western blotting. B : Transfected HCEnC-21T cells were seeded to 100% confluence on an electric cell–substrate impedance sensing (ECIS) electrode array. A high current (arrow) was applied to the circular electrode underneath the central region of the cell monolayer, thus clearing the circular area. Migration of cells into the cleared area was monitored for 12 h after wounding by measuring impedance (ohms) across the electrode at 4,000 Hz. Three wells for each condition were measured; n = 3. Error bars = standard error of the mean (SEM).

Article Snippet: 8W1E electric cell–substrate impedance sensing (ECIS) Cultureware™ disposable electrode arrays (Applied BioPhysics, Troy, NY) were stabilized with F99 medium according to the manufacturer’s protocol and then were coated with 40 μg/cm 2 chondroitin sulfate A (Sigma-Aldrich) and 400 ng/cm 2 laminin (Sigma-Aldrich) in PBS for 2 h. Twenty-four hours after transfection, the HCEnC-21T cells were reseeded at 100% confluency in the stabilized electrode arrays.

Techniques: Migration, Transfection, Western Blot, Electric Cell-substrate Impedance Sensing

Journal: Molecular Vision

Article Title: Elucidating the molecular basis of PPCD: Effects of decreased ZEB1 expression on corneal endothelial cell function

doi:

Figure Lengend Snippet: The impact of ZEB1 reduction on cell barrier function in in HCEnC-21T cells. A : HCEnC-21T cells were transfected with ZEB1 siRNA-C or scrambled siRNA (sc-siRNA), and knockdown of ZEB1 was confirmed with western blotting. B : Impedance (ohm) at 4,000 Hz, interendothelial resistance (Rb), cell attachment resistance (alpha), and plasma membrane capacitance (Cm) were measured for 48 h after seeding on electric cell–substrate impedance sensing (ECIS) electrode arrays. Three wells for each condition were measured; n = 3. Error bars = standard error of the mean (SEM). Statistical significance (p<0.05) is noted for a range when two or more sequential time points demonstrated statistical significance.

Article Snippet: 8W1E electric cell–substrate impedance sensing (ECIS) Cultureware™ disposable electrode arrays (Applied BioPhysics, Troy, NY) were stabilized with F99 medium according to the manufacturer’s protocol and then were coated with 40 μg/cm 2 chondroitin sulfate A (Sigma-Aldrich) and 400 ng/cm 2 laminin (Sigma-Aldrich) in PBS for 2 h. Twenty-four hours after transfection, the HCEnC-21T cells were reseeded at 100% confluency in the stabilized electrode arrays.

Techniques: Transfection, Western Blot, Cell Attachment Assay, Electric Cell-substrate Impedance Sensing