design: Triplicate 25 mL cultures of strains indicated above were grown in 7H9-ADS until they reached an OD600 of 0.3 to 0.5. Cultures were then treated with the indicated concentration of TTM, 100nM Q203, or a DMSO vehicle control, for 5 hrs. Following a duplicate wash with Chelex treated PBS Tween-80, cells were collected by centrifugation (3,700  g, 10 min), suspended, and resuspended in 1 mL of TRIzol reagent. Cells in 1 mL of TRIzol were mechanically disrupted with zirconia beads via 2x45 sec pulses in a BioSpec Mini24 BeadBeater with 5 min intervening rest periods on ice. Following lysis, beads were removed by centrifugation (20,000  g, 5 min). After RiboGreen quantification and quality control by Agilent BioAnalyzer, 500 ng of total RNA underwent ribosomal depletion with the NEBNext rRNA Depletion Kit (Bacteria) (NEB catalog #E7850) and library preparation with the TruSeq Stranded Total RNA LT Kit (Illumina catalog #RS-122-1202) according to instructions provided by the manufacturer with 8 cycles of PCR. Samples were barcoded and run on a NovaSeq 6000 in a PE100 run, using the NovaSeq 6000 S4 Reagent Kit (200 Cycles) (Illumina). On average, 27 million paired reads were generated per sample. Post-run demultiplexing and adapter removal were performed and fastq files were inspected using fastqc. Trimmed fastq files were then aligned to the reference genome (M. tuberculosis H37Rv; NC_000962.3) using bwa mem. Bam files were sorted and merged using samtools and gene counts were obtained using featureCounts from the Bioconductor Rsubread package. Differentially expressed genes were identified using the DESeq2 R package and subsequent analysis of gene expression was performed.