Mechanical Stretching of HTM Cells

HTM cells were cultured in collagen-coated plates (Flexcell International, Burlington, VT, USA, BioFlex plates, BF-3001C), and after 90% confluency, the plates were fixed on a FlexCell (Flexcell International, FX-6000 Tension System). The cells were stretched at 0.69 Hz frequency with 15% stretching for 8 h for gene expression analysis and 24 h for protein expression analysis based on previously published reports [65 (link),66 (link),67 (link),68 (link)]. Cells and media were collected and processed, as mentioned later, for RNA analysis and immunoblotting.

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Soundararajan A., Ghag S.A., Vuda S.S., Wang T, & Pattabiraman P.P. (2021). Cathepsin K Regulates Intraocular Pressure by Modulating Extracellular Matrix Remodeling and Actin-Bundling in the Trabecular Meshwork Outflow Pathway. Cells, 10(11), 2864.

Publication 2021

FX-6000 Tension System

Cells Collagen Gene expression analysis Protein

Corresponding Organization : Neurosciences Institute

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Variable analysis

independent variables

Stretching frequency (0.69 Hz)
Stretching magnitude (15%)

dependent variables

Gene expression
Protein expression

control variables

Cell type (HTM cells)
Culturing conditions (collagen-coated plates)
Time points (8 h for gene expression, 24 h for protein expression)

controls

No controls explicitly mentioned.

Annotations

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