multi copy plasmid pmv261 expressing mtb dnae1d228n m tuberculosis strain h37rv h37rv wild type mtb strain h37rv  (ATCC)

Journal: Molecular microbiology

Article Title: Chemical activation of adenylyl cyclase Rv1625c inhibits growth of Mycobacterium tuberculosis on cholesterol and modulates intramacrophage signaling

doi: 10.1111/mmi.13701

Figure Lengend Snippet: H37Rv was grown for 5 d in 7H12 media with 0.2 % acetate and 100 μM cholesterol, and then washed in 7H12 media to remove extracellular cAMP. The bacterial cells were resuspended in 7H12 0.2 % acetate and 100 μM cholesterol, with either 0.1 % DMSO (black) or 10 μM V-58 (gray). Aliquots of the cultures were removed at the indicated times and processed to quantify cAMP concentrations which were normalized to 108 CFU. A) Amount of cAMP found in the A) cytoplasmic fraction; and B) extracellular media (secreted fraction). The data shown are the means of two biologically independent experiments with the error bars as the standard deviation between experiments. *p≤0.05, **p≤0.005 as determined by unpaired two tailed t-test.

Article Snippet: Maintenance of bacterial strains and cell cultures Wild-type Mtb H37Rv (ATCC 25618), CDC1551, M. bovis BCG Pasteur and Msm mc 2 155 were used where indicated.

Techniques: Standard Deviation, Two Tailed Test

Journal: Molecular microbiology

Article Title: Chemical activation of adenylyl cyclase Rv1625c inhibits growth of Mycobacterium tuberculosis on cholesterol and modulates intramacrophage signaling

doi: 10.1111/mmi.13701

Figure Lengend Snippet: J774.16 murine macrophages were infected with live Mtb H37Rv or CDC1551, and heat-killed CDC1551 were treated with either 0.1 % DMSO (black bars) or 10 μM V-58 (gray bars). cAMP levels were quantified by RIA from the A) intracellular bacteria and B) macrophage lysates. The data shown are the means of two biologically independent experiments. Error bars represent the standard deviation between replicates. C) Macrophages were infected with Mtb CDC1551 at the indicated MOI and treated with 10 ng/mL LPS, 250 μM 8-Br-cAMP or V-58 at the indicated concentrations for 18 h and supernatant TNF-α levels were determined by ELISA. Data are means of four biologically independent experiments and the error bars represent the standard deviation between experiments. Statistical significance was determined with a two-tailed unpaired t-test, *p≤0.05, **p≤0.005 and ****p≤0.0001.

Article Snippet: Maintenance of bacterial strains and cell cultures Wild-type Mtb H37Rv (ATCC 25618), CDC1551, M. bovis BCG Pasteur and Msm mc 2 155 were used where indicated.

Techniques: Infection, Bacteria, Standard Deviation, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Molecular microbiology

Article Title: Chemical activation of adenylyl cyclase Rv1625c inhibits growth of Mycobacterium tuberculosis on cholesterol and modulates intramacrophage signaling

doi: 10.1111/mmi.13701

Figure Lengend Snippet: Mtb H37Rv was grown in either A) Mycomedia, B) 7H12 100 μM cholesterol, C) 7H12 20 mM propionate or D) 7H12 100 μM cholesterol and 20 mM acetate in the presence of either 0.1 % DMSO (circles) or 10 μM V-58 (open squares). The OD620 of each culture was measured at the indicated time points. Show are the means of two independent experiments. Statistical significance was determined with a two-tailed unpaired t-test, *p≤0.05, **p≤0.005

Article Snippet: Maintenance of bacterial strains and cell cultures Wild-type Mtb H37Rv (ATCC 25618), CDC1551, M. bovis BCG Pasteur and Msm mc 2 155 were used where indicated.

Techniques: Two Tailed Test

Journal: Molecular microbiology

Article Title: Chemical activation of adenylyl cyclase Rv1625c inhibits growth of Mycobacterium tuberculosis on cholesterol and modulates intramacrophage signaling

doi: 10.1111/mmi.13701

Figure Lengend Snippet: , Mtb H37Rv was grown in either Mycomedia, 7H12 100 μM cholesterol, 7H12 20 mM propionate or 7H12 100 μM cholesterol and 20 mM acetate in the presence of either 0.1 % DMSO (black) or 10 μM V-58 (gray). cAMP levels were quantified by RIA at 7 d for each culture and normalized to 108 CFU, with bars representing the total cAMP produced (sum of cAMP in the bacterial lysate and extracellular medium). Data shown are means of two biologically independent experiments and the error bars represent the standard deviation. Statistical significance was determined with a two-tailed unpaired t-test, *p≤0.05, **p≤0.005.

Article Snippet: Maintenance of bacterial strains and cell cultures Wild-type Mtb H37Rv (ATCC 25618), CDC1551, M. bovis BCG Pasteur and Msm mc 2 155 were used where indicated.

Techniques: Produced, Standard Deviation, Two Tailed Test