Immortalized cell lines HAP 1 and MEF were used for holographic imaging assay. The cells were cultured as described previously (6, 7) . Briefly, HAP 1 cells were grown Iscove's Modified Dulbecco's Medium (IMDM), supplemented with 10% FBS, 10 mL per L of Antibiotic Antimycotic Solution Penicillin/Streptomycin/Amphoterichin B; Sigma Aldrich) and 2 mM L-Glutamine. MEF cells were grown in high glucose Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS, 10 mL per L of Antibiotic Antimycotic Solution Penicillin/Streptomycin/Amphoterichin B (Sigma Aldrich), 2 mM L-Glutamine, non-essential amino acids. Both cell lines were maintained in a humidified cell incubator, at 37°C under a 5% CO2 atmosphere. HAP 1 Δ (c+δ) cell line that lacks c and δ-subunits of ATP synthase were used for the study of the role of ATP synthase in high-conductance PTP.
Antibiotic antimycotic solution penicillin streptomycin amphoterichin b
Manufactured by Merck Group
Antibiotic Antimycotic Solution Penicillin/Streptomycin/Amphoterichin B is a sterile, liquid solution containing the antibiotics penicillin, streptomycin, and the antifungal agent amphotericin B. This solution is commonly used as a supplement in cell culture media to prevent bacterial and fungal contamination.
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2 protocols using antibiotic antimycotic solution penicillin streptomycin amphoterichin b
1
Imaging Immortalized Cell Lines for ATP Synthase
2
Immortalized Cell Lines for ATP Synthase and ANT Studies
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Immortalized HAP 1 and MEF cell lines were used for this study.6 (link),8 (link) HAP 1 cells were grown Iscove’s Modified Dulbecco’s Medium (IMDM), supplemented with 10% Heat-Inactivated Fetal Bovine Serum (HI FBS; Life Technologies), 10 mL per L of Antibiotic Antimycotic Solution (Penicillin/Streptomycin/Amphoterichin B; Sigma Aldrich) and 2 mM L-Glutamine. MEF cells were grown in high-glucose Dulbecco’s Modified Eagle Medium (DMEM; Cytiva) supplemented with 10% HI FBS, 10 mL per L of Antibiotic Antimycotic Solution, and 1X Non-Essential Amino acids (NEAA; Lonza). HAP 1 Δ (c+δ) cell line that lacks c and δ-subunits of ATP synthase was used for the study of the role of ATP synthase in high-conductance PTP. MEF ANT Triple KO cell line lacking 3 ANT genes was used to study the role of ANT in high conductance PTP. MEF ANT Triple KO cells were grown in the same media as MEF WT cells with the addition of 1mM Sodium Pyruvate (Gibco) and 25mg/500mL Uridine (Sigma Aldrich). Cells were maintained in a humidified cell incubator, at 37°C under a 5% CO2 atmosphere.
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