Fig. 4: sAEL057 treatment leads to altered cholesterol metabolism in Mtb.

a, b Mtb was pre-grown for 6 days in cholesterol-supplemented media prior to the addition of sAEL057 at 10 µM or DMSO control. [4-¹⁴C] radiolabeled cholesterol was added at 24 h posttreatment and ¹⁴CO₂ was measured 6 h later. n = 6 from two replicate experiments. c, d The bacterial reporter, prpD’::GFP smyc’::mCherry, was used to assess levels of propionyl-CoA with sAEL057 treatment in cholesterol-supplemented media. c % GFP-positive bacteria were determined by gating on mCherry positive bacteria and measuring the GFP-positive signal using a BD Symphony analyzer. d Bacteria positive for both mCherry and GFP fluorescence were analyzed to determine the GFP median fluorescence intensity (MFI). n = 2 from replicate experiments. Statistical significance was assessed using a student’s unpaired t-test. Error bars indicate standard deviation. Gating strategy displayed in Supplemental Fig. 7. Source data and P values for all main figures available in Supplementary Data 1.