Fig. 5. LytB-Bs and LytC-Bs can exogenously cleave CW of foreign species.
a CWs were isolated from Bs (PY79), Bm (OS2), or Sa (MRSA), labeled with RBB, mixed with purified LytB-Bs, LytC-Bs or both. The cleaved products were separated by centrifugation and absorbance of supernatant was measured at 595 nm. Shown are mean ± SEM of at least three independent experiments. b Bs sacculi derived from PY79 cells were incubated for 30 min with purified LytB-Bs, LytC-Bs or both, spotted onto poly l-lysine coated coverslips and visualized by XHR-SEM. Scale bars represent 500 nm. c Bs (PY79), Bm (OS2) or Sa (MRSA) cells were incubated for 10 min with purified LytB-Bs, LytC-Bs or both, stained with PI and visualized by fluorescence microscopy. Shown are overlay images of phase contrast (gray) and fluorescence from PI staining (red). Scale bar represents 5 μm. d Quantitation of PI uptake by Bs (PY79), Bm (OS2), or Sa (MRSA) cells treated as described in c. Shown is the percentage of PI-labeled cells and mean ± SEM of at least three independent experiments (ncells = 400). NT indicates no treatment (samples contained only buffer). The experiments were repeated at least three times independently with similar results. Source data are provided as a Source Data file.