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I-confluence [3] determines whether a conflicting operation will preserve application invariants based on application dependent correctness criterion.
Is the confluence of these two movements a mere historical accident?
MVECs transduced with DN or WT calpain-I or empty vector were grown to confluence in full medium.
Tenocytes were grown to confluence in an eight-well glass slide (Lab-Tek®II Chamber Slide™ System Nalge Nunc International Corp.., Rochester, NY, USA).
Total RNA was isolated from recombinant clones grown to confluence in a 96-well plate using ZR-96 mini RNA isolation I kit (Zymo Research).
In our study we seeded the cell cultures overnight after the cells had reached 80% confluence (i.e. proliferative phase) to ensure the cells were expressing the maximum amount of σ2 receptors in the cells, also to optimise signal-to-noise ratio in cell uptake studies.
Cells were subcultured to a fresh culture dish when growth reached 70 90% confluence, i.e. every 2 3 days.
The highest DON level (4000 ng/mL) resulted in a TEER-decrease even below the level of cellular confluence, i.e. 1 kOhm, after 72 hours exposure.
The N/C ratio decreased to 1/1 when cells reached confluence, i.e. when most cells were in the G0/G1 phase of the cell cycle, suggesting that cells need a reduced nuclear environment to proliferate.
Cell density (i.e. confluence) was calculated using the IncuCyte software.
All experiments showed optimal confluence (i.e., > 70%) except one, which displayed a confluence of about 50 60%.
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