Cells in all four flasks were grown simultaneously at 37°C, 200 rpm, until the most concentrated culture grew to an OD600 of 1.0 (Flask D).
Approximately 1 × 10 cells/T25 flasks were grown for 1 day before adding garlic or inhibitors.
OVCAR-3 cells (3 × 10) seeded in 75 cm2 flasks were grown for 72 h at 37°C to 80% confluency.
Cells in T-150 flasks were grown in a 37 °C incubator (Thermo Scientific, Waltham, MA, USA) provided with 7% carbon dioxide air.
The cells in the remaining flasks were grown and passaged to allow examination of effects in distant progeny according to the methods described by Seymour et al (1986).
Cultures (volume, 175 ml in 1 L flasks) were grown at 37°C, under shaking conditions (200 rpm) until the OD600nm reached 0.2.
A single colony was used to inoculate an initial 50 mL preculture in a 250 mL glass Erlenmeyer flask, which was grown for 1 day at 20 °C with shaking at 200 rpm.
For anaerobic expression, an overnight culture was used to inoculate (1% vol/vol) 1 L of medium in a 2.8 L baffled Fernbach flask, and the culture was grown at 37°C shaking at 150 rpm under N2 for 3 5 hours to an OD600nm ∼0.8.
For shaking flask cultivation, C. glutamicum was grown in the complex medium LB (lysogeny broth) or in the chemically defined medium CGXII [ 98], but containing only 2% glucose (instead of 4%) and 30 mg l-1 instead of 0.03 mg l-1 protocatechuic acid, in 250 ml shaking flasks at 30°C and 300 rpm.
A single 175 cm flask of RK-13 cells was grown to confluence (defined as a continuous cell monolayer) and split into three separate flasks.
