ATP flux was examined by luminometry, as previously described5 (link),46 (link). Primary calvarial cells isolated from Panx3 KO mice were seeded at 104 cells/well in a 96-well plate, and induced by osteogenic culture media for 14 days. The cells were then washed with PBS, followed by incubation in PBS for 2 min. The supernatant was collected and assayed with luciferase and luciferin (Promega). The luminescence was measured using an Infinite 200 PRO (TECAN).
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Quantifying Osteogenic ATP Flux
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Corresponding Organization : National Cancer Institute
Other organizations : University of Pennsylvania
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Variable analysis
independent variables
- Panx3 KO (Knockout) mice
- ATP flux
- Cell seeding density (10^4 cells/well)
- Osteogenic culture media
- Incubation time (14 days)
- Incubation in PBS (2 min)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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