Figure 2.
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.

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.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal