Fig. 5

Fig. 5 SMAD3-VEGF interactions influence blood-brain-barrier integrity in a zebrafish amyloidosis model.

a We pharmacologically treated transgenic zebrafish model Tg(kdrl:GFP) with vegfr2 blockers to reduce VEGF signaling. b Double immunostaining for GFP and pSMAD3 coupled to DAPI nuclear counterstain. Lower panels indicate percentage of pSMAD3⁺ cells. In endothelia panel, insets indicate neuronal pSMAD3. Inside brackets the number of analyzed cells are shown. Vegfr2 blockage increased the percentage of pSMAD3⁺ endothelial cells and pericytes (GFP⁺/DAPI⁺). c pERK and GFP double immunostaining coupled to DAPI nuclear counterstain in control and vegfr2 blocker treated zebrafish models. Inside brackets the number of analyzed spots are shown. Vegfr2 blocking decreased pERK/GFP colocalization in zebrafish models. d Double immunostaining for ZO-1 and GFP in control and vegfr2 blocker-treated zebrafish models coupled to DAPI nuclear counterstain. Treatment caused decreased colocalization of ZO-1/GFP, indicating impaired integrity in zebrafish brain vasculature. Correlation graph between random measurements between pERK/GFP vs ZO-1/GFP indicated strong association. R indicates the correlation coefficient. Scale bars equal 5 µm (b) and 10 µm (c,d).e Colocalization coefficients of pERK/GFP and ZO-1/GFP are correlated. With decreased pERK, ZO-1 also reduces, similarly, high pERK expressing vascular cells also has high ZO-1 levels. GFP always marks the vasculature, and therefore is common to both separate correlations. Source data are provided as a Source Data file. Figure 5/panel a Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs license.