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. 2023 Jul 11;14:4123. doi: 10.1038/s41467-023-39723-8

Fig. 3. Dysfunction of lipid II synthesis associates with an mbl mutant lethality.

Fig. 3

a Growth curves during murGB repression. B. subtilis strain YK1540 (Pspac-murG-murB) was cultured in NB containing 0.1 mM IPTG (blue line) at 37 °C. The exponentially growing cells were diluted into fresh NB (without IPTG, red line) with 10 μg/ml MC (purple line) or 10 mM Mg2+ (orange line) and cultured at 37 °C for OD600 measurements. The means were obtained from three independent experiments. Error bars indicate standard deviation (SD). Source data are provided as a Source Data file. b Effects of MC on lethal effect and cell morphology during murGB repression. Cell morphologies of about 200 cells under each condition shown in panel a were classified into three types (rod, sphere/bulging or phase pale) based on the phase contrast images as shown in panel c and Suppremenratly a. The results were obtained from three independent experiments. Error bars indicate SD. Source data are provided as a Source Data file. c Phase contrast micrographs of Pspac-murG-murB (YK1540) cells were captured in the course of carrying out the growth curves shown in panel a. Scale bars represent 5 μm. d Effects of murGB expression levels on mbl mutant growth. YK1540 (Pspac-murG-murB) and YK2665 (Δmbl Pspac-murG-murB) strains were streaked on NA plates containing various concentrations of IPTG, with or without 10 μg/ml MC or 10 mM Mg2+. The plates were incubated for 18 h at 37 °C. e Effects of murGB expression levels on cell morphology of an mbl mutant. Phase contrast micrographs of strain YK2665 (Δmbl Pspac-murG-murB) in liquid NB containing 10 mM Mg2+, with 0.05 mM or 1 mM IPTG (+Mg2+). The exponentially growing cells were diluted into fresh NB (no added Mg2+) with 0.05 or 1 mM IPTG and incubated for 2–3 h (No add). Scale bars represent 5 μm. f Growth rescue of an mbl mutant by murG overexpression. YK2700 (Δmbl amyE::Pxyl- murB) and YK2701 (Δmbl amyE::Pxyl- murG) were streaked on NA plates with or without 10 mM Mg2+ or 0.5% xylose, and incubated for 18 h at 37 °C. The figures are representative of at least three independent experiments.