Fig. 3

Morphologies of mamA depleted cells and rescue by deletion of MSMEG_3214. a Microscopy of the mamA KD strain treated with ATc for 12 h. Cells were stained with SYTO 24 (green) and FM 4-64FX (red) to visualize DNA and membranes, respectively. The images represent cells of three major phenotypic groups observed. The left column shows DNA stained only, the center column shows membrane stained only, and the right column shows the merged fluorescent images. b Strains with depleted mamA (mamA KD), nonspecific CRISPRi control with an sgRNA sequence that does not target any specific part of M. smegmatis genome (Nonspecific sgRNA), mamA KD in an MSMEG_3214-deleted background (mamA KD ΔMSMEG_3214), and the ΔMSMEG_3214 strain with a nonspecific sgRNA (Nonspecific sgRNA ΔMSMEG_3214) were incubated in 7H9 in presence (+) or in absence of ATc to activate or not the CRISPRi knockdown system, respectively