Fig. 2: Cl-amidine reduced endothelial activation, EndMT and proliferation in CCM.
a, Representative images of the cerebellum of Ccm3-iECKO vehicle-treated mice (upper panel) and Cl-amidine-treated (10 mg kg−1 d−1) mice (lower panel) stained with DAPI (blue), ICAM-1 (red) and ILB4 (white). Insets with white line highlight magnifications to the right. b,c, Quantification of total ICAM-1+ area (b) and vascular (ILB4+) ICAM-1+ area (c) in the cerebellum of Ccm3-iECKO vehicle-treated (n = 13) and Cl-amidine-treated (10 mg kg−1 d−1; n = 8) mice. d, Representative images of endothelial cells undergoing EndMT in the cerebellum of Ccm3-iECKO mice stained with DAPI (blue), Sca1 (yellow) and podocalyxin (magenta). Co-localization of Sca1 and podocalyxin is shown as white. Insets with dashed, white line are shown in the magnifications to the right. Insets with white line are shown in the magnifications to the bottom, showing zoomed-in areas with EndMT with nuclei stained with DAPI (blue). e, Representative images of endothelial cells undergoing EndMT in the cerebellum of Ccm3-iECKO mice stained with DAPI (blue), vimentin (yellow) and ILB4 (magenta). Co-localization of vimentin and ILB4 markers is shown as white. Insets with dashed, white line are shown in the magnifications to the right. Insets with white line are shown in the magnifications to the bottom, showing zoomed-in areas with EndMT with nuclei stained with DAPI (blue). f, Quantification of vascular (podocalyxin+) Sca1+ area in the cerebellum of Ccm3-iECKO vehicle-treated (n = 13) and Cl-amidine-treated (10 mg kg−1 d−1; n = 8) mice. g, Quantification of vascular (podocalyxin+) vimentin+ area in the cerebellum of Ccm3-iECKO vehicle-treated (n = 12) and Cl-amidine-treated (10 mg kg−1 d−1; n = 8) mice. h, Representative images of the cerebellum of Ccm3-iECKO vehicle-treated mice (upper panel) and Cl-amidine-treated (10 mg kg−1 d−1) mice (lower panel) stained with Ki67 (magenta), ERG (green) and podocalyxin (white). Insets with dashed, white lines are shown in the magnifications to the right. i, Quantification of Ki67+ vascular (ERG+) area in the cerebellum of Ccm3-iECKO vehicle-treated (n = 13) and Cl-amidine-treated (10 mg kg−1 d−1; n = 8) mice. In the graphs, each data point represents one biological replicate; the bar indicates the median of each group; and the error bars represent the IQR. Statistical significance was determined using a Mann–Whitney U-test (two-tailed). cb, cerebellum.
