Figure 2.

Characterization of EbpC_fm surface expression by TX82 and its ebpABC_fm deletion and complementation derivates. (A) Surface expression of EbpC_fm in different growth stages and growth media, using whole-cell ELISA. Cells were washed and adjusted to OD_{600 nm} = 0.5 before coating on wells. EbpC_fm expression was detected using an affinity-purified anti-EbpC_fm antibody.³⁰ Bars represent the means of absorbance at 590 nm ± SD from four independent assays, each performed in triplicate. (B) Flow cytometry analysis. Labeling by the anti-EbpC_fm antibody is shown for each strain and by a pre-immune antibody (PI)³⁰ for TX82, and indicated as log fluorescence intensity on the X-axis. Bacteria were analyzed using side scatter as the threshold for detection. Each histogram represents 50,000 events of bacterium-sized particles. (C) Western blot of mutanolysin extracts. Five µg of mutanolysin cell wall extracts were electrophoresed and blotted onto the membrane and probed with affinity-purified anti-EbpC_fm antibodies or pre-immune antibodies. Lane 1, TX82; lane 2, TX82ΔebpABC_fm; lane 3, TX82ΔebpABC_fm (pAT392); lane 4, TX82ΔebpABC_fm (pAT392::ebpABC_fm), lane 5, rec-EbpC_fm.³⁰