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Figure 8 Combination of bezafibrate and urolithin A restores normal phenotype in Costello zebrafish. Phenotype analysis of (A) control zebrafish or (B–H) Costello zebrafish injected with HRASV12 plasmid at 5 dpf, after 4 days of treatment with (B) DMSO, (C) bezafibrate, (D) urolithin A, or (E and F) bezafibrate (BZ) and urolithin A (UA) in combination. Costello zebrafish developed edema (B–D, arrows) or hemorrhages and vascularization defects (B and D, asterisks). (F) Treatment with a combination of 10 μM BZ and 5 μM UA markedly reduced the percentage of abnormal embryos. (G) Illustration of the phenotypes observed in Costello zebrafish at 5 dpf after expression of HRASV12 plasmid. (H) Expression level of the mitochondrial protein TOM20 in control or HRAS p.G12V embryos. TOM20 protein content determined by Western blot was normalized to the total protein content. (I) Survival rate of Costello zebrafish after 4 days of treatment with DMSO, bezafibrate, or UA alone or in combination. Data were normalized to day 1 of treatment. (J) Percentage of defect appearance in Costello zebrafish after 4 days of treatment with DMSO, bezafibrate, or UA alone or in combination. Data are expressed as number of embryos or as the mean value of TOM20 expression. Unpaired t test was used to compare the 2 groups of zebrafish (WT or Costello). *P < 0.05.