Figure 3
In vitro functional analysis of MaMIF. (A) Data shows the MaMIF-tautomerase activity under the increasing concentration of this protein. (B) Graphical representation showing the influence of increasing concentrations of dopachrome on MIF-tautomerization activity. (C) Determination of the Km and Vmax for the substrate-dependent MIF-tautomerase activity. (D) Figure shows the expression of Tnf-α, Il-6, Il-1β, and Vegf in PBMCs following stimulation with BSA (100 ng/ml), MaMIF (100 ng/ml), Polymyxin B (30 μg/ml), MaMIF + Polymyxin B, ISO-1 (100 μg/ml) and MaMIF + ISO-1 for 4 hours (n = 3). Data represents the mean ± SEM, *p < 0.05; **p < 0.001; ***p < 0.0001 using one-way ANOVA with Bonferroni’s Multiple Comparison Test. (E) Results of the trans-well migration assay show the effect of MaMIF on the migration property of PBMCs. Digital images of the stained trans-well porous membrane show PBMCs (blue colored) that have migrated through the pores after treatment with BSA (100 ng/ml), MaMIF (100 ng/ml), Polymyxin B (30 μg/ml), MaMIF + Polymyxin B, ISO-1 (100 μg/ml) and MaMIF + ISO-1 for 2 hours. Quantification of the number of migrated cells in different conditions is shown through the bar graph (n = 3). Each bar represents the mean ± SEM, ***p < 0.0001 using a Student t-test. The scale bar is 100 µm.
