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. 2021 May 25;118(22):e2104166118. doi: 10.1073/pnas.2104166118

Fig. 1.

Fig. 1.

Serum fractions B11 and C3 inhibit IT4var19 IE binding to brain endothelial cells. (A) Chromatographic fractionation of serum by size exclusion chromatography. IgG-depleted serum was run on a Superdex 200 Increase 10/300 GL column. Protein fractions were eluted using a linear gradient and the absorbance (mAbs) monitored at 280 nm. The arrows show the binding inhibitory fractions B11 and C3. (B) Binding of IT4var19 IEs to HBEC-5i in binding medium with 10% human serum, IgG-depleted human serum, or serum fractions (A6-C6). A no-serum control was included. Each data point is from two independent experiments done in duplicate, and the mean and SEM are shown for each fraction. Fractions B11 and C3 inhibited the binding the most. These fractions were run on MS/MS, and the components were run to determine their effect on binding as shown in C to identify potential inhibitors for binding. (C) Binding of IT4var19 IEs to HBEC-5i in binding medium with proteins identified by MS/MS in the serum inhibitory B11 and C3 fractions (attractin, C1s, C4b, factor H, IgD, ITIH1, ITIH2, and kappa light chain). Data are mean ± SEM, and each data point is from an independent experiment. **P < 0.01; ***P < 0.001.