Fig 5. Inducing cAMP synthesis independent of V-59 and Rv1625c is sufficient to block cholesterol utilization.

(A) Total cAMP induced in TetOn-cAMP Mtb. Cultures were treated with V-59 (10 μΜ), Atc (500 ng/mL or 50 ng/mL), or EtOH and samples were collected after 24 hours. Data are normalized as total cAMP per 10⁸ Mtb and are from two experiments with two technical replicates each. (B) Impact of inducing TetOn-cAMP on the growth of Mtb in cholesterol media. Cultures were treated with V-59 (10 μM) or Atc for the duration of the experiment. Data are from two experiments with three technical replicates. (C) Catabolic release of ¹⁴CO₂ from [4-¹⁴C]-cholesterol in the TetOn-cAMP strain treated with V-59, Atc, or EtOH. Data are from two experiments with three technical replicates, normalized to OD and quantified relative to EtOH EtOH is the vehicle control throughout. All data are means ± SD (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, One-way ANOVA with Dunnet’s multiple comparisons test).