FIG 3.

Rv1625c is required for GSK286-dependent growth inhibition and block of cholesterol metabolism in M. tuberculosis. (A) MIC curves for GSK286 in M. tuberculosis grown in cholesterol media. Inhibition was calculated using alamarBlue fluorescence with 100% inhibition established using rifampicin treatment (10 μM). All of the inhibition values are normalized to this value. The x-axis values are logarithms of concentrations expressed as a molar unit. (B) GFP fluorescence inhibition curves for GSK286 using the prpD′::GFP reporter in cholesterol-acetate media. The strains included WT (wild-type M. tuberculosis), 2×Rv1625c (WT strain carrying a plasmid expressing rv1625c from an hsp60 promoter), and TnRv1625c (a transposon-disrupted mutant with a nonfunctional Rv1625c). Each point in the fluorescence inhibition curve was obtained by quantifying the GFP signal from 10,000 bacteria, and error bars indicate means ± the SD (n = 4) from two independent replicates with two technical replicates for each.