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Cultures were maintained in phenol red free Neurobasal medium with 2 mM glutamine, N-2 supplement, and gentamycin (Invitrogen).
Experiments were performed at room temperature during which cells were maintained in phenol red-free DMEM supplemented with 10% FCS.
CaOV-3 cells are routinely maintained in phenol red-positive RPMI 1640 medium supplemented with 10% FBS (v/v), 2 mM glutamine and 50 µg/ml gentamicin.
To establish an androgen-independent cell line, LNCaP was first maintained in phenol red-free DMEM/F-12 medium (Gibco) with 10% charcoal/dextran-treated FBS (Biowest).
LNCaP C-33 cells are androgen-sensitive and routinely maintained in phenol red-positive RPMI 1640 medium supplemented with 5% FBS (v/v), 2 mM glutamine and 50 µg/ml gentamicin [28].
MCF-7 cells were maintained in phenol free EMEM/CD-FBS media for 4 days prior to drug treatments (in triplicate) and MCF-7/shRNA ERRα RNAi cells were maintained in normal media containing whole serum (described above).
OVCAR-3 cells are routinely maintained in phenol red-positive RPMI 1640 medium supplemented with 20% FBS (v/v), 2 mM glutamine, pancreatic bovine insulin (10 µg/ml) and gentamicin (50 µg/ml).
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Cells were maintained in phenol-red free DME containing 10% fetal calf serum and 50 mM Hepes or EBSS where indicated for live-cell imaging experiments.
Cells maintained in phenol-red-free DMEM (Gibcol-BRL, USA) with 2.5% dextran charcoal-stripped fetal calf serum (Biochrom AG, Germany) for 48 72 h were switched to medium without 12 h before treatment with the agents indicated.
For estrogen-deprivation conditions, MCF7 cells were maintained in phenol-red-free DMEM (Invitrogen) containing 5% charcoal-stripped FBS (HyClone).
TAM-R and LTED were maintained in phenol-red free DMEM containing 10% charcoal stripped fetal calf serum (SFCS).
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