Fig. 2: INU1-fab causes cerebral thrombosis.
a,b, Horizontal sections of murine brains (scale 50 µm) taken 20 min after vehicle (control; n = 6), INU1-IgG (0.75 µg g−1; n = 7) or INU1-fab (0.5 µg g−1; n = 7 and 6, respectively) i.v. injection reveal cerebral thrombi predominantly located in large vessels in between the two hemispheres (upper panels), or at the outside of the cortex (lower panels). Thrombi were visualized using H&E (a) or anti-GPIX (p0p615; green counterstained with anti-CD31, magenta and 4,6-diamidino-2-phenylindole, gray) on cryo-sections (Cryo) (b). c, A cranial window was mounted on top of the superior sagittal sinus and blood flow and thrombus formation were monitored using intravital confocal microscopy. Platelets were stained using an anti-GPIX derivative15 (green), the endothelial lining was stained with anti-CD31 and the vessel lumen with fluorescently labeled bovine serum albumin (both depicted in magenta); scale 100 µm. Shown are snapshots from intravital microscopy videos (Supplementary Videos 2–4) at the indicated time points after i.v. injection of vehicle (control; n = 3), INU1-IgG (0.75 µg g−1; n = 4) or INU1-fab (0.5 µg g−1; n = 6). d, Three-dimensional reconstruction of LSFM images of brains from mice that were perfusion-fixed 20 min after i.v. application of vehicle (control; n = 4), INU1-IgG (0.75 µg g−1; n = 5) or INU1-fab (0.5 µg g−1; n = 4). Platelets (anti-GPIX derivative15, green) and endothelial cells (anti-CD31, anti-CD105, both in magenta) were stained in vivo, paraformaldehyde-fixed brains were cleared using benzyl alcohol/benzyl benzoate and imaged on a custom-built light-sheet fluorescence microscope15; the scale is indicated as a grid with a size of 500 µm (Supplementary Videos 5–8). Numerical data are displayed as box whisker blots with median being displayed as central line and whiskers indicating minimum and maximum.
