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. 2017 Jul 19;7:326. doi: 10.3389/fcimb.2017.00326

Figure 2.

Figure 2

Interaction of FimH adhesin with porcine calreticulin. (A) Far-Western blotting analysis of FimH adhesin binding to recombinant porcine CRT. CRT (0.5 μg) was subjected to SDS–PAGE and transferred onto nitrocellulose. CFimH, C63FimH and EFimH were incubated with CRT immobilized on the membrane and then detected with anti-FimH rabbit polyclonal antibody and secondary anti-rabbit antibody. (B) Interaction of CFimH, C63FimH and EFimH with immobilized CRT analyzed with an SPR. Lines represent different concentrations of analyte (0.5, 1, 2, 4, and 8 μM) in HBS buffer. Binding data were collected at a flow rate of 30 μl/min. (C) Dissociation constants of scFimH adhesins determined by SPR analysis. (D) Adherence of SCΔfimH/C to IPEC-J2 cells preincubated for 1 h with PBS buffer (control), CFimH adhesin, C63FimH adhesin or EFimH adhesin. Data are means ± SD in triplicate assays, which were independently repeated three times.