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HL3-containing medium at varying concentrations of extracellular Ca+ were used (0.375 mM for mEJP recordings and 1.5 mM for EJP recordings).
The cells were immediately counted and resuspended in serum-free L15 medium at varying concentrations (for instance, 20,000, 10,000 or 5,000 cells in 10 μl, the injection volume).
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Medium was replaced with medium containing drugs at varying concentrations, and the cells were then incubated at 37 °C for 24 h before 10 μl of CCK-8 (Dojindo Laboratories, Rockville, MA, USA) was added to each well.
Then, the cell culture medium was replaced by normal DMEM medium containing lunasin at varying concentrations (0, 10, 50, 100, and 200 μM).
Sfib were stimulated with 10 ng/ml TNF-α and pretreated for 2 hours with medium or MTX at varying doses (0, 0.01, 0.1, 1, 10 μM) in the presence or absence of adenosine deaminase, DPCPX (adenosine A1 receptor antagonist) or DMPX (adenosine A2 receptor antagonist).
SB715992 was then applied directly to RPMI 1640 medium in experimental cultures at varying concentration.
A total of 300 μl of incubation medium containing radioactively labeled ligand proteins at varying concentrations was added and the cells were incubated for 3 h at 4°C.
To address this issue, the cells were seeded at varying densities and the medium was changed to KO/KR on day 2 of plating.
In a recent paper by Suggett et al. (2008) H2O2 production (and release to the medium) by two Symbiodinium types was measured at varying temperature and light intensities [41].
In all intermediate cases, the effect of the medium on the detected signal produces fluctuations of at varying, due to the roughly linear expected dependence of on the modulation frequency.
The extracts (20 μl/well) at varying concentrations, diluted in culture medium, were added in duplicate.
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