Proliferation of ASM cells based on DNA content was measured as described previously (70 (link)). Briefly, ASM cells were grown in 96-well plates (~5,000 cells/ well), serum-deprived, transfected with siRNA, exposed to 1% CSE for 48h and incubated with CyQuant NF (Invitrogen) for 30 min. Dye (CyQuant) binding to DNA (fluorescence) was measured on a FlexStation3 microplate reader (Molecular Devices, Sunnyvale, CA). Dye calibrations and normalization were performed, as previously described (70 (link)).
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Quantifying ASM Cell Proliferation via DNA
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Corresponding Organization : Mayo Clinic
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Variable analysis
independent variables
- SiRNA transfection
- Exposure to 1% CSE for 48h
- Proliferation of ASM cells based on DNA content
- ASM cells grown in 96-well plates (~5,000 cells/well)
- Serum-deprived ASM cells
- Incubation with CyQuant NF (Invitrogen) for 30 min
- Dye (CyQuant) binding to DNA (fluorescence) measured on a FlexStation3 microplate reader (Molecular Devices, Sunnyvale, CA)
- Dye calibrations and normalization performed as previously described (70 (link))
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