Promotion of S. aureus biofilm formation by exogenously supplemented recombinant proteins.
Single colonies of S. aureus MS4 were grown in 5 ml BHI medium at 37°C overnight. The overnight cultures were 1,000-fold diluted in 5 ml BHIG medium mentedupples with the indicated concentrations (0.05 to 1 M) of purified Eap and Efb, and the diluted cultures were then placed into 96-well polystyrene plates. The plates were incubated at 37°C for 24 h under static conditions. After the incubation, the plates were gently shaken and the culture media were discarded. The plates were then washed twice with PBS buffer and dried under atmosphere. The remaining biofilmswere stainedwith 0.1%crystal violet, and the absorbance at 595 nm was monitored using a microplate reader (Tecan). Assays were performed in triplicate.