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10 μl of media from each well was moved to a new 96-well plate and 100 μl of LDH reaction mix was added to each well.
An amount of 50 μl from each extract was filter sterilized and added to the seeded cells (after removal of 50 μl of cell culture media from each well).
Following treatment, media from each compartment were used for quantification of U. parvum (quantitative PCR), interleukin (IL -8 (enzyme-lIL -8 immunosorbent assay), and matrix metalloproteinase (MMP)-2 and MMP-9 actimmunosorbentassay.
The international press would be present as well as the media from each of the finalist cities.
Cells lysates and conditioned media from each situation were collected and stored for immunoblotting, as described [44].
We also cultured embryonic wildtype C57Bl/6 or NSE-Db thalami and harvested the conditioned media from each.
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Bio-media from each unit in the series of submerged biofilters were cut into small pieces, and the microbial film was suspended in phosphate-buffered saline solution.
We next examined the effect of the pknG deletion on GABA production from glucose, using C. glutamicum strains GAD, GADΔpknG, and W. As glucose was consumed, GABA formation was detected in the media from cultures of each during stationary phase (Figure 2A).
We therefore generated a 'balanced dataset' that includes the same number of media sources from each political leaning (four of each are selected randomly).
Each experimental condition was assayed in duplicate and duplicate media samples from each were measured using a Cortisol EIA (DSL-10-200) from Diagnostic Systems Laboratories (Webster, TX), according to the manufacturer's directions.
Relative cytotoxicity measurements were determined by subtracting the background (1% FBS media only) from each experimental sample.
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