Correlation between induced TdmhMtb activity and increased non-selective permeability of mycobacterial envelope under limiting nutrients. (A) Influx of glycerol over two hours in mc27000 incubated in normal (7H9TyOADC) and nutrient-limiting media (7H9Ty) for indicated time. (B) Regeneration of Mtb(Erd) in 7H9Ty with 0.1% v/v glycerol from a culture maintained in 7H9Ty for indicated time. (C) Accumulation of acetate, phosphorus and glycerol over 30 minutes by mc2155:ΔtdmhMs transformed with either pJL37, or pRv3451. (D) Accumulation of acetate, phosphorus and glycerol over two hours by cultures of mc27000, mc27000:ΔtdmhMtb, and mc27000:ΔtdmhMtb expressing Rv3451 (mc27000:ΔtdmhMtb:p51), Rv3452 (mc27000:ΔtdmhMtb:p52), or Rv3451-52 (mc27000:ΔtdmhMtb:p5152) in nutrient-limiting media (also see Fig. S3). (E) Growth of Mtb(Erd), ΔtdmhMtb mutant, and the complemented strain (mc27000:ΔtdmhMtb:p5152) in 7H9Ty with 0.1% v/v glycerol. (F) Sensitivity of 4-week biofilms of Mtb (Erd), Mtb (Erd):ΔtdmhMtb, and Mtb (Erd):ΔtdmhMtb:p5152 to H2O2 (40mM) and lysozyme (6μg/mL). (Figure 3, related to Figure S3)