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When the OD600 of the cultured medium reached 0.6, the culture was induced with isopropyl-β-d-thiogalactopyranoside (IPTG) at a final concentration of 1 mM for 4 h.
After treatment, cultures were washed with fresh cultured medium and grown for the next 24 hours.
Based on the TLC analysis data, the cultured medium and cell-free culture supernatant of L. plantarum LG42 cells contained GABA, but not the cytoplasmic fraction.
The cultured medium was used to inoculate the fermentation culture.
The cultured medium was natural seawater enriched with MES medium.
For analysis of IL-2 in a cultured medium, the supernatants were analysed with a Mouse IL-2 ELISA Ready-SET-Go!
EN-RAGE protein was detected in the cultured medium and increased significantly by the addition of IL-6.
Before the reagent was added, 3.0 mL of cultured medium was transferred to other dishes at 37 °C, and the membrane was transferred to each medium in turn (1 mL reagent medium for 30 min; 1 mL wash medium for 10 min; and 1 mL remaining medium for monitoring of bioluminescence).
The concentration of NO in the cultured medium was measured indirectly by analysis of nitrite levels using the Griess reaction.
A positive control was cultured without Hg amendment, while other controls were alum salts with Hg-amended medium without yeast inoculation, and cultured medium without alum treatment.
Acetylsalicylic acid (ASA) was purchased from sigma (Madrid, Spain), and dissolved in the cell cultured medium.
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