<p>A. S. <i>aureus</i> Newman overnight cultured cells were washed in water, 5 M NaCl, 2% CHAPS, 8 M urea, 6 M guanidine HCl, or 3 M LiCl. In another sample, <i>S</i>. <i>aureus</i> cells were digested with lysostaphin and treated with 2% CHAPS. Samples were centrifuged and the amount of PSMα3 or PSMα1+δ-toxin in the supernatant was measured by HPLC. Vertical axis represents the amounts of PSM recovered from <i>S</i>. <i>aureus</i> cells (1.33 ml bacterial culture). Data are means±standard errors from three independent experiments. B. The centrifuged supernatants obtained in <i>A</i> were analyzed by SDS-PAGE. Proteins in the supernatants were precipitated with 10% trichloroacetic acid and electrophoresed on a 12.5% SDS polyacrylamide gel. The gel was stained by Coomassie brilliant blue. Each lane contains proteins from the same number of <i>S</i>. <i>aureus</i> cells (0.09 ml bacterial culture). C. <i>S</i>. <i>aureus</i> Newman overnight cultured cells were washed in 6 M guanidine HCl or 2% SDS. Samples were centrifuged and the amount of PSMα3 in the supernatant was measured by HPLC. Vertical axis represents the amount of PSMα3 recovered from <i>S</i>. <i>aureus</i> cells (1.33 ml bacterial culture). Data are means±standard errors from triplicate experiments.</p
