Mycobacterium susceptibility to ivermectin by inhibition of eccD3, an ESX-3 secretion system component.
Fig 6
Characterization of CRISPRi M. smegmatis strains.
(a) Representative image of M. smegmatis strains cultures (PLJR962-eccD3-gRNA, PLJR962-control-gRNA and PLJR962-mmpL3-gRNA) grown in 7H10 medium supplemented with OADC, kanamycin 0.03 µM (20 μg/mL) and without or with ATc. (b) A significant difference in growth was observed in M. smegmatis PLJR962-mmpL3-gRNA strain without or with ATc, using Student’s t-test (p= 0.0168) bars represent the average of the replicates and error bars illustrate standard deviation of the data. (c) Relative expression of eccD3 gene, average of technical triplicates of each strain without (-) and with (+) ATc (anhydrotetracycline) 0.0002 μM (100 ng/mL). For (b) and (c) eccD3 corresponds to M. smegmatis PLJR962-eccD3-gRNA strain, ctrl corresponds to M. smegmatis PLJR962-control-gRNA strain, WT corresponds to M. smegmatis wild type strain and mmpL3 corresponds to M. smegmatis PLJR962-mmpL3-gRNA. The rod with the asterisk shows significant differences between the relative expression of eccD3 gene in M. smegmatis PLJR962-eccD3-gRNA strain using Student’s t-test (p=0.0062).