Fig. 5
Immunohistochemistry, histology, immunofluorescence and autoradiography of LJP1586-treated fMOG-EAE rat brain sections with supporting quantitative data. (a, h) OX-42 immunohistochemical staining of activated microglia (the contours define the drawn ROIs used for quantification). (b, i) H&E staining. (c, j) Anti-VAP-1 immunofluorescence staining with secondary antibody captured with Panoramic MIDI scanner. (d, k) VAP-1 targeted 68Ga-DOTA-Siglec-9 ex vivo autoradiography. Autoradiography brain sections are outlined according to the corresponding sections in immunohistochemistry. Low power scale bars = 2 mm. High power (e, l) OX-42, (f, m) H&E, (g, n) anti-VAP-1 immunofluorescence staining with secondary antibody captured with confocal microscope. Tissue auto-fluorescence signal (red colour) is separated from specific VAP-1 staining (green) using the spectral imaging followed by linear un-mixing based on the reference spectra.. High power scale bars = 50 μm. Red arrows point to VAP-1 positive vessels in brain vasculature. Quantification of autoradiography data (o) and area of activated microglia (p) for LJP1586 treated fMOG-EAE. *P < 0.05, **P < 0.01, ***P < 0.001