To examine the effect of EZH2 inhibition on gel contraction, we followed the procedure as described (25 (link)). SSc dermal fibroblasts were treated with GSK126 (0.5–10 µM) for 72 h before suspension in culture media at 2 × 106 cells/mL. Cells were then mixed with collagen solution from the Cell Contraction Assay kit (Cell Biolabs) and plated in a 24-well plate. Culture media was added after the collagen polymerized. After 1 d, the collagen matrix was released, and the size of the collagen gel was measured and analyzed after 5 h using ImageJ (24 (link)).
Protocol detail
Evaluating EZH2 Inhibition on Fibroblast Contraction
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Other organizations : University of Michigan–Ann Arbor
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Variable analysis
independent variables
- GSK126 concentration (0.5–10 µM)
- Size of the collagen gel
- Cell density (2 × 10^6 cells/mL)
- Cell type (SSc dermal fibroblasts)
- Culture duration (72 h for treatment, 5 h for gel contraction measurement)
- No positive controls mentioned
- No negative controls mentioned
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