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. 2013 May 6;2:10.3402/jev.v2i0.20181. doi: 10.3402/jev.v2i0.20181

Fig. 3.

Fig. 3

Antibodies raised against rHtrA-OMV recognize C. muridarum HtrA and bind the EB surface. (A) HtrA-specific ELISA IgG titres. Sera from mice immunized with different OMV formulations (CM rHtrA-OMV, empty OMV and empty OMV mixed with rHtrA) and with purified rHtrA (doses of 20 and 1 µg) were collected 2 weeks after the third immunization and IgG titres were analyzed by ELISA. Bars represent the mean titres of each mouse group expressed as the reciprocal of the serum dilution yielding an OD value of 0.5. (B) Western blot analysis of CM EBs. Total extracts of CM EBs and purified CM rHtrA were loaded on the gel, transferred to nitrocellulose and immunoblots were probed using antibodies raised against CM rHtrA-OMV (left panel) and empty OMV (right panel). Molecular weight standards are reported on the left of each panel. (C) Flow cytometry analysis of CM EBs. CM EBs were incubated with anti-CM rHtrA-OMV (open peak) and anti-empty OMV antibodies (shaded peak) and surface-bound antibodies were detected by flow cytometer using a R-phycoerythrin-conjugated secondary antibody.