Supplementary Figure 8: Rescue of the M. smegmatis ∆qcrCAB strain by the qcrCAB operon in the plasmid and verification of expression.

a. Colonies of ∆qcrCAB strain harboring the construct PST-KT^qcrCAB grow significantly larger than those of the control strain (PST-KT). The 7H9 agar plates are shown after 5 days of growth at 37 °C. b. 7H9 liquid culture. PST-KT^qcrCAB bearing cells show comparable growth to wild-type M. smegmatis (bearing PST-KT) while control plasmid (PST-KT) bearing ∆qcrCAB cells show impaired growth. The OD₆₀₀ after 48 hr of growth at 37°C (starting OD = 0.05) are shown. Error bars indicate SD (n=3). c. Verification of the expression of SC proteins. Detection of FLAG tagged cytochrome b in Ms∆qcrCAB+PST-KT^qcrCAB strain by western blotting. A single band is detected at 57 kDa. Lanes 1 and 2 are loaded with lysates from Ms∆qcrCAB+PST-KT^qcrCAB and WT+PST-KT cells, respectively. d. Detection of cytochrome cc by heme staining. Membranes purified from the ∆qcrCAB+PST-KT^qcrCAB were resolved on SDS-PAGE and subjected to heme staining. The molecular weight of marker proteins is shown on the left.