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. 2012 Jul 27;7(7):e40411. doi: 10.1371/journal.pone.0040411

Figure 5. Activity of rSpyCEP and PMN transmigration.

Figure 5

(A) Comparison of rSpyCEP and native SpyCEP activity on IL-8. SDS-PAGE (18% Tris-Glycine) and silver staining after 2 h of digestion with 5 ng, 1 ng, 0.2 ng, 0.04 ng or 0 ng (ctr lane) of rSpyCEP+3348ΔspyCEP extracts (left). Cell wall extracts containing comparable amounts of native SpyCEP from 3348 extracts (right). Full and trace arrowheads indicate intact and cleaved IL-8, respectively. Lower panel: control Western blot showing relative SpyCEP amounts compared to 5 ng of rSpyCEP (ctr). (B,C) SpyCEP effect on PMN transmigration. (B) Murine or (C) human PMN migration in response to KC or IL-8 (white), respectively, in the presence of rSpyCEP (black) or rSpyCEP* (grey). (D) Counteraction of rSpyCEP activity by specific antibodies. Human PMN migration using IL-8 (white); IL-8, rSpyCEP and α-SpyCEP (black); and IL-8, rSpyCEP, α-Spy0269 (grey). Data represent means plus SEM of one representative experiment using triplicates. Statistical significance was tested by unpaired Student T, (*) P<0.05, (**) P<0.01, (***), P<0.001.