Fig. 5

IKK-inhibiting viral degradation of Foxj1 and CSF/brain barrier disruption. a IHC staining of primary EC cultures without (Ctrl) or with indicated treatment conditions, labeled with Foxj1 and VP16 antibodies, and DAPI. Cultures were treated with viruses for 16 h. Quantifications showing Foxj1⁺ cells per total DAPI nuclei in treatment conditions as percentage of Ctrl condition in each experiment (read dashed line). * P < 0.03, Wilcoxon two-sample test, n = 4, z = 1.225. Scale bar: 15 µm. b IHC staining of ependymal wholemounts from P28 FOXJ1-GFP animals injected with PBS (Ctrl, left panels) or HSV-1 (right panels), labeled with GFP and acetylated tubulin (A-tub) antibodies. Samples were harvested 3 days after injection. Note the large GFP⁺ ependymal gaps in HSV-1 injected condition, showing ventricular wall breakdown exposing underlying brain parenchyma (A-tub⁺ cellular patches). Scale bar: 50 µm. c Representative IHC staining of ependymal wholemounts from P28 animals treated with PBS (Ctrl), HSV-1, or HSV-1 + MLN4924, labeled with VP16 antibody showing infected areas (VP16⁺). Samples were harvested 2 days after treatments. Scale bar: 50 µm. d DAPI IHC staining of ependymal wholemounts, comparing HSV-1 treated or HSV-1 + MLN4924 co-treated conditions. Image stacks represent the top 20 µm from ependymal surface. Dashed lines indicate areas of ventricular disruption where cell patches are absent from the surface. Quantifications represent average number of cellular gaps observed during each condition. * P < 0.0004, Wilcoxon two-sample test, n = 6, z = 1.988. Scale bar: 25 µm. Box plots show mean (+), median (−), quartiles (boxes), range (whiskers)