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In Fig. 3, we find different evolutionary tracks depending on τSF in the right-hand panel, whereas these tracks, with the metallicity normalized to the critical metallicity, are well overlaid on each other in the left-hand panel.
Cells were seeded onto Matrigel-coated 6-well plates (1.5 million cells per well) overlaid 3 hours later with a second layer of Matrigel (ECM-sandwich) or onto a single ALI-3D membrane in each well of a 6-well plate.
The reasoning for the phone-numbers-like array beneath the regular A, B, and C buttons which irritatingly run backwards, from the perspective of a SEGA user was that an overlay could be, well, overlaid atop it, explaining the function of each button dependent on the game.
The reasoning for the phone-numbers-like array beneath the regular A, B and C buttons – which irritatingly run backwards, from the perspective of a SEGA user – was that an overlay could be, well, overlaid atop it, explaining the function of each button dependent on the game.
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The culture in each well was overlaid with 50 µl mineral oil prior to starting the assays.
After infection, each well was overlaid with culture medium containing 0, 0.01, 0.1, 1.0, and 10.0 U/ml of penicillin (Sigma-Aldrich) respectively.
The next day, each well was overlaid with 300 microliters of collagen-I at a concentration of 0.5 mg/ml in serum-free medium together with inhibitors as indicated.
Each well was overlaid with a 6 μl layer of light mineral oil (Sigma) to minimize evaporation.
After the incubation, each well was overlaid with approx. 10 BHK-21 cells and the plates were incubated for five days at 37°C in 5% CO2.
After 10 min, the incubation buffer was removed and transferred into a counting vial; the cells remaining in the well were overlaid with a 1% SDS solution, thereby disintegrated and the resulting solution transferred into a counting vial.
Next, wells were overlaid with 5×103 cells per well in a 0.3% agar.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com