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. 2003 Jan;71(1):242–253. doi: 10.1128/IAI.71.1.242-253.2003

FIG. 5.

FIG. 5.

Lack of activity of YspB-(His)6 or YspC secreted by the Ysc injectisome of Y. enterocolitica ΔHOPEM. (A) Bacteria (2.5 × 107; C) or the culture supernatant of 5 × 109 bacteria (S) was loaded on an SDS-polyacrylamide gel and analyzed by Western blotting with an anti-His (C-terminal) monoclonal antibody or an anti-YspC polyclonal antibody. The detection was performed with Supersignal chemiluminescent substrate (Pierce). (B) BCECF release from preloaded macrophages upon infection with Y. enterocolitica ΔHOPEM, ΔHOPEMBD, ΔHOPEMBD(pMRS74), and ΔHOPEMBD(pBF15). Results are expressed as the percentages of Triton-lysed macrophages after subtraction of the value measured in the supernatants of uninfected cells. Data show the means and standard deviations of assays performed in triplicate. (C) SDS-polyacrylamide gel stained with Coomassie blue loaded with the culture supernatant of 5 × 109 Y. enterocolitica ΔHOPEM, ΔHOPEMBD, and ΔHOPEMBD(pBF15) bacteria expressing sycB, yspB, and yspC from the yopE promoter under in vitro conditions. MW, molecular weights in thousands.