Fig 3. Spatial distribution of Sbp in S. epidermidis cultures.

(A) Preparations of cell wall associated proteins and 10-fold concentrated supernatants from S. epidermidis 1457 (biofilm-positive) and 1585 (biofilm-negative) after static overnight growth were separated by SDS PAGE and blotted onto PVDF-membranes. Sbp was detected after incubation with rabbit anti-rSbp antiserum and anti-rabbit IgG coupled to peroxidase by chemiluminescence. (B) Growth phase dependent regulation of Sbp. Cell wall associated proteins were prepared from S. epidermidis 1457, 1457ΔsarA, and 1457ΔrnaIII at different time points during adherent growth in TSB. At each time point cell numbers were adjusted to an identical A₆₀₀ before cell surface associated proteins were isolated by boiling in LDS buffer. After separation of surface associated proteins by SDS-PAGE and blotting onto a PVDF membrane Sbp was detected by chemiluminescence using a rabbit anti-rSbp antiserum and a peroxidase-coupled anti-rabbit IgG. SDS-PAGE analysis proved loading of gels with similar total protein amounts (S2 Fig.).