To determine the potentiating effects of 2,2′-bipyridyl (BP) (Sigma-Aldrich, Burlington, MA, USA) on the bactericidal activity of SA23, mid-log-phase wild-type and katG loss-of-function mutant (11 (link)) M. bovis BCG cultures were diluted at an OD600 value of 0.05 in catalase-free complete 7H9 broth. The cell suspensions were then treated with test compound SA23 at 0.5× and 1× MIC90 at 37°C and 110 rpm for 5 days with or without the presence of 80 μM BP. After 5 days of drug treatment, aliquots of the cultures were serially diluted and plated on complete 7H10 agar for CFU counting. Each experiment was independently repeated thrice, with one representative set of results (containing two technical replicates) being shown in Fig. 3 .
2 2 bipyridyl bp
Manufactured by Merck Group
Sourced in United States
2,2'-bipyridyl (BP) is a heterocyclic organic compound with the chemical formula C10H8N2. It is a white crystalline solid that serves as a ligand in coordination chemistry and as a precursor in organic synthesis.
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2 protocols using 2 2 bipyridyl bp
1
Assessing Potentiating Effects of 2,2'-Bipyridyl on Mycobacterial Killing
2
Stabilizing HIF-α and Studying FGF Signaling in HeLa Cells
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HeLa cells (ATCC CCL--2 TM ) were cultured in DMEM supplemented with 10% heat--inactivated fetal bovine serum (FBS), 100 U/ml penicillin, 0.1 mg/ml streptomycin and 2 mM L--glutamine (Life Technologies). Treatments were performed on 70 % confluent cells after overnight starvation in the absence of FBS. To stabilize HIFα subunits, 20 mM diethyl succinate (DES) or 0.1 mM 2,2' -bipyridyl (BP) (Sigma) was added and cells further incubated in serum--free medium for approximately 16--h.
In some experiments, 100 ng/ml of recombinant human fibroblast growth factor -acidic (FGF) (Life technology), was used in serum--free medium for 30 min and cells collected after 24--h in complete medium.
GPx8 silenced HeLa cells contained a vector for stable expression of small interfering RNA addressed to GPx8 (SilenciX technology) and were purchased from Tebu--bio, which also provided HeLa cells transfected with a control shRNA. The percentage of silencing was 97%, as quantified by the manufacturer. GPx8 silenced cells and controls thereof were grown as above except that the medium contained 4 mM L--glutamine, 110 mg/l sodium pyruvate and 125 µg/ml hygromycin B (Life Technologies). These were treated with 100 ng/ml of FGF or 20 µg/ml insulin for 10 min in serum free medium and lysed as described below. Proteins were quantified by the Bradford reagent (Sigma).
In some experiments, 100 ng/ml of recombinant human fibroblast growth factor -acidic (FGF) (Life technology), was used in serum--free medium for 30 min and cells collected after 24--h in complete medium.
GPx8 silenced HeLa cells contained a vector for stable expression of small interfering RNA addressed to GPx8 (SilenciX technology) and were purchased from Tebu--bio, which also provided HeLa cells transfected with a control shRNA. The percentage of silencing was 97%, as quantified by the manufacturer. GPx8 silenced cells and controls thereof were grown as above except that the medium contained 4 mM L--glutamine, 110 mg/l sodium pyruvate and 125 µg/ml hygromycin B (Life Technologies). These were treated with 100 ng/ml of FGF or 20 µg/ml insulin for 10 min in serum free medium and lysed as described below. Proteins were quantified by the Bradford reagent (Sigma).
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