Vented flasks

The bacterial strains, plasmids, and primers used in this work are listed in Table 1. Mycobacterium tuberculosis strains were grown in 7H9 liquid media (Difco) supplemented with 0.2% glycerol, 0.05% Tween 80, 0.5% bovine serum albumin (BSA), 0.2% dextrose, and 0.085% sodium chloride (ADN) (referred to as “7H9c” from here on) or Sauton minimal media (3.7 mM potassium phosphate, monobasic; 2.4 mM magnesium sulfate; 30 mM L-asparagine; 3.5 mM zinc sulfate; 9.5 mM citric acid; 6.0% glycerol; 0.005% ferric ammonium citrate; 0.05% Tween-80). Cultures were grown at 37°C without agitation in vented flasks (Corning). For M. tuberculosis growth on solid media, Middlebrook 7H11 agar (Difco and Remel) was supplemented with Middlebrook OADC (oleic acid, albumin, dextrose, and catalase; BBL). Mycobacterium tuberculosis strains were grown in 50 µg/mL kanamycin, or 50 µg/mL hygromycin when necessary.
For CuSO₄ solutions, stock solutions were made by dissolving the appropriate amount of CuSO₄ powder (Fisher Scientific) in water and filter-sterilizing with a 0.45-µm filter. For aldehyde solutions, 50 mM pHBA was made with pHBA powder dissolved in water and filter-sterilized using a 0.45-µm filter. MG (5.5 M) was diluted in sterile water just before use. Both aldehydes were purchased from Sigma-Aldrich, Inc.

T. brucei brucei bloodstream form (s427) was cultured in vitro in HMI11 [73 (link)] and CMM [44 (link)] supplemented with 10% fetal calf serum (FCS) Gold (PAA, Piscataway, NJ) at 37°C, 5% CO₂. Initial 5 ml cultures in 25 ml vented flasks (Corning) were grown to a maximum density of 4 × 10⁶ cells/ml and subcultured by 1-in-100 or 1-in-1,000 dilution every 2 or 3 days, respectively. A hemocytometer (Neubauer) was used for cell counts.