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Fig. 7
CL-387,785 can effectively reduce the levels of C99 and AICD without perturbing Notch-dependent somite development in a zebrafish model of amyloidopathy. (A) Schematic presentation of the DNA construct encoding recombinant C99-GFP-P2A-GFP. CI, chimeric intron; CMV, human cytomegalovirus promoter; P2A, porcine teschovirus 2A peptide. (B) Zebrafish embryos at the one-cell stage were injected with C99-GFP-P2A-GFP plasmid and treated with the indicated compounds from 48 h postfertilization (hpf) to 72 hpf at 28 °C. Clarified lysates derived from treated embryos were analyzed by Western blotting with anti-GFP (for C99, C83, AICD, and P2A-GFP plasmid injection control), anti-A?1?16 (6E10, for full-length C99), and anti-GAPDH. The asterisk indicates unidentified bands. WT, wild type. (C) Phenotypes of C99-expressing zebrafish embryos treated with the indicated compounds were imaged by bright-field microscopy and fluorescence microscopy. The number of embryos exhibiting a curved trunk at 72 hpf was recorded (n = 90).