Fig 7. CisS supports cell morphology defects and lack of srtA and htrA leads to antibiotic susceptibility.

(A) Statistical analysis of pili expressing cells labeled with α-EbpC immune serum and Alexa Fluor 568 secondary antibody. Results are represented as bar graphs with standard error of mean. Combined data from three independent experiments were shown. Statistical analysis was performed by the 2-way ANOVA and Tukey multiple comparison test using GraphPad. *** P ≤ 0.001; **** P ≤ 0.0001; P > 0.05, differences not significant (ns). (B) The cell wall was stained with Vancomycin-FL conjugate (green). DAPI (blue) was used to visualize DNA. Representative images were shown. Scale bars represent 1 μm (C) The maximum cell width of the various mutant strains was measured and quantified using MicrobeJ plugin in ImageJ and are represented as violin plot. Cells that were not in-phase were excluded from the analyses. A total of at least 900 cells were sampled per strain. Dashed line represents quartiles; horizontal solid line represents the median (D) Minimal inhibitory concentration of various antibiotics for E. faecalis mutants. ^# indicates the median MICs reported from 2 biological replicates. ATCC 29212 is the positive control strain that fits the standard MIC of gentamicin (4–16 μg/mL) (E) List of selected genes relevant to this study and their log₂-fold change in transcription in OG1RF WT treated with subinhibitory concentrations of daptomycin. Differentially expressed genes are highlighted in bold font and yellow background.