Fig. 3

SDS-PAGE (a) Coomassie brilliant blue staining and Western blotting analysis of recombinant fusion proteins Flg-HA2–2-4M2ehs, FlgSh-HA2–2-4M2ehs by anti-M2e mAb 14C2 (b) and anti-Flg mAb 93,713 (c); immunostaining with HRP-conjugated second antibodies and TMB substrate. Positions of molecular weight markers (MM) are indicated. 1 - Flg-H2–2-4M2ehs; 2 – FlgSh-H2–2-4M2ehs. d Response of HEK-Blue™ hTLR5 cells to recombinant proteins. HEK-Blue™ hTLR5 and HEK-Blue™ Null1 (control) cells were incubated in HEK-Blue™ Detection medium and stimulated with 100 ng/ml recFLA-ST (positive control), 100 μg/ml ODN2006 (negative control) and 100 ng/ml of FliC, Flg-HA2–2-4M2ehs, or FlgSh-HA2–2-4M2ehs. After 24 h incubation, the levels of NF-kB-induced SEAP were determined by reading the OD at 655 nm. Error bars represent the s.d. obtained from three biological replicates