Curative effects upon Talarozole treatment of krit1 mutant zebrafish and retinoic acid treatment of siCCM2 depleted HUVECs. (a) Treatment scheme for zebrafish from 18 to 48 hpf using Talarozole. Endocardial cell numbers (a readout for CCM-associated cardiac phenotype in zebrafish) were analyzed after treatment in two independent experiments. (b) Representative images of zebrafish heart morphologies visualized by the transgene kdrl:EGFP at 48 hpf after treatment. Upper: heart morphology of controls treated with DMSO. Lower: hearts treated with Talarozole. Scale bars: 100 µm. (c) Endocardial cell counts at 48 hpf of zebrafish treated with several doses of Talarozole or DMSO (*P < 0.05; ****P < 0.0001 vs. control). Each data point represents one heart. (d) Treatment regimen of HUVECS. SiRNA transfection was done for 24 h, followed by treatment with RA 24 h later. Cell morphologies were analyzed after 48 h of incubation and antibody staining. (e) Immunohistochemistry of HUVECS for beta-catenin, F-actin, and DAPI. First column: control cells (siCT) with wild-type CCM2 expression. Second column: untreated siCCM2-depleted HUVECs. Third column: representative image showing siCCM2 HUVECs treated with 100 nM RA. Scale bars: 10 µm.
