Fig. 4

Seaporter TLR5 cell line responses to flagellin and SAPN. SeaPorter TLR5 cell-line was exposed to varying concentrations of each indicated protein, and the level of TLR5 stimulation was determined by the level of SEAP expression. Fold increase in SEAP expression for each protein sample over non-treated controls, error bars are standard error of the means. A two-way ANOVA model was fit with protein concentration and type as factors. There was a significant protein concentration by type interaction (p < 0.001); this indicated that the differences across types depended on the concentration and that the differences across concentrations varied by type. Specifically, there weren’t statistically significant differences across types at the two lowest concentrations (0.01 and 0.1), but there were significant differences between types at the 1, 10, 100, and 1000 ng/mL concentrations (p < 0.001 for all). Subsequent pairwise contrasts at these 4 concentrations found that the 5A11 Restricted CD8 + group (the recombinant protein without flagellin) was significantly different from the other 3 groups in all cases except for the 5A11 Restricted CD8 + vs. CD8-SAPN comparison at the 1 ng/mL concentration. In addition, at each of these 4 concentrations, the Empty-SAPN was significantly different (greater than) from the CD8-SAPN. There was a significant concentration effect for all protein types (p < 0.001) except 5A11 Restricted CD8 + (p > 0.9)