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Fig 3
A Domains of M. abscessus LeuRS. Zinc domain, pink; editing domain, cyan; catalytic domain, grey; leucine-rich domain, green; anticodon-binding domain, purple; c-terminus, yellow. B Amino acid alignment of part of the editing domain of LeuRS M. abscessus ATCC 19977, EPT mutant, and reference mycobacteria. Non-consensus residues are in red. Genbank accession numbers: P9WFV1.1 (M. tuberculosis) 11896261 (M. avium) 6224275 (M. marinum). C Dose-response curves of M. abscessus ATCC 19977 (black circles), D436H mutant (red squares), M. abscessus empty vector (white diamonds), M. abscessus pMVhsp60_leuS (pink triangles) and M. abscessus pMV306hsp60_leuSD436H (crosses). Data shown is mean ± SD from one experiment representative of two. D leuS is an essential gene in M. abscessus using CRISPRi gene knockdown. M. abscessus carrying integrated empty CRISPRi vector (EV, circles), CRISPRi:mmpL4b as a non-essential gene control (squares), or CRISPRi:leuS (triangles) were grown with/without 0.01 ?g/mL ATc. Data is mean ± SD from biological triplicates. E MIC99 to EPT and RFB in CRISPRi knockdown of leuS in M. abscessus. **, p = 0.005; ns, no statistical significance by one-way ANOVA with Dunnett?s multiple comparisons test. Data shown is mean ± SD from three biological replicates.