Humoral immune response in mice immunized with mTLNE. (A) The immunoreactivity of serum from mice immunized with mTLNE was determined by Western blotting. The total proteins of EV71 were harvested from infected cell lysate, and the cell lysate from the uninfected RD cells was used as control. Lanes 1: uninfected RD cell lysate. Lanes 2: the total proteins of EV71. The position of the three capsid proteins of EV71 (VP0, VP1 and VP2) was indicated by arrow, respectively. (B) The lgG antibody titer against EV71 was measured by IFA at two weeks after each immunization. (C and D) Neutralization antibody titer against EV71 strain AH08/06 (genotype C4) and prototype strain BrCr (genotype A) were measured by microneutralization assay at two weeks after the final immunization. Dotted lines represent limits of detection.