CCM pathology promotes the expression of genes and proteins related to the coagulation cascade and hypoxia in vivo. (A) Visium spatial transcriptomics illustrating the expression of F3 in wild-type and Ccm3-iECKO P8 cerebellum sections. (B) Quantification of F3 in the cerebellum (cb) of wild-type and Ccm3-iECKO mice. (C) Visium spatial transcriptomics illustrating the expression of Serpine1 in wild-type and Ccm3-iECKO cerebellums. (D) Quantification of Serpine1 in the cerebellum (cb) of wild-type and Ccm3-iECKO mice. (E) Visium spatial transcriptomics illustrating the expression of Vegfa in wild-type and Ccm3-iECKO cerebellums. (F) Quantification of Vegfa in the cerebellum (cb) of wild-type and Ccm3-iECKO mice. (G) Hematoxylin and eosin-stained Ccm3-iECKO P8 cerebellum sections with lesions outlined in green. (H) RNAscope images of representative wild-type (right panel) and Ccm3-iECKO (left panel) cerebellum sections; a region is highlighted with a white box and magnified to the right. Lesions are outlined in dotted white lines and regions of interest are marked with a white asterisk; 4′,6-diamidino-2-phenylindole(DAPI) (blue), Cldn5 (green), F3 (magenta). (I) Quantification of F3 in the cerebellum (cb) of wild-type and Ccm3-iECKO mice (P = .0635). (J) RNAscope images of representative wild-type (right panel) and Ccm3-iECKO (left panel) cerebellum sections. Lesions are outlined in dotted white lines; DAPI (blue), Cldn5 (green), Serpine1 (magenta). (K) Quantification of Serpine1+Cldn5+ endothelial cells in the cerebellum (cb) of wild-type and Ccm3-iECKO mice (P = .0159). (L) RNAscope images of representative wild-type (right panel) and Ccm3-iECKO (left panel) cerebellum sections; a region is highlighted with a white box and magnified to the right. Lesions are outlined in dotted white lines; DAPI (blue), Cldn5 (green), Vegfa (magenta). (M) Quantification of Vegfa in the cerebellum (cb) of wild-type and Ccm3-iECKO mice (P = .0635). (N) Primary human brain endothelial cells (HBECs) transduced with shRNA (scramble or CCM3) blotted and quantified (O) CCM3 (P = .0035), (P) TF (P = .3650), (Q) VEGF-A (P = .8346), and (R) PAI-1 (P = .0178). In the graphs N-R, n = 3 independent experiments. In graphs B, D, F, I, K, and M each data point represents 1 biological replicate (n = 2-8 mice per group), the bar indicates the mean of each group, and the error bars represent the standard deviation. For the RNAscope graphs, a Mann-Whitney U test was used to compare wild-type mice with Ccm3-iECKO mice. For the western blot quantifications, a t-test was used to compare shScramble with shCCM3 cells. The corresponding P values are indicated on each graph. L, lesion.