Fig. 5

siRNA mediated RFC1 knockdown led to inner BRB breakdown and endogenous IgG extravasation. The wholemount retinas were labeled by Lectin (cyan) and incubated with Cy3 labeled goat anti-mouse antibody (yellow) to visualize the extravasated endogenous mouse IgG. A RFC1-siRNA mediated knockdown led to significant IgG extravasation (red arrows) in microvessels while scrambled-siRNA treated retinas showed no extravascular IgG signal (p = 0.001; n = 3). B 1 h ischemia led to microvessel constrictions (white arrows) but no IgG extravasation. However, knocking down RFC1 before ischemia provoked inner BRB disruption leading to endogenous IgG extravasation (red arrows) (p = 0.05; n = 3). Nuclei were labelled with Hoechst 33258 (blue) in all images. Scale bars= 10 μm