Figure 5

(A) Pericardial injection of PC inhibitor. Confocal images of fli1a:EGFP-labeled cartilage structures show PCI injection improves chondrocyte morphology, organization, and differentiation in pmm2^m/m embryos. White boxes, central portion of Meckel’s cartilage; orange boxes, lateral regions evaluated for phenotypic rescue. Scale bars: 10 μm. Percentage values indicate number of scored embryos exhibiting pictured phenotype. n = 30 embryos over 4 experiments. (B) Parameters assessed for rescue of central portion of Meckel’s cartilage (white boxes). Cells were scored for percentage of intercalated, ratio between long and short axis (measure of roundness), and percentage of cells with vacuoles. n = 8–15 treated embryos over 4 experiments. (C) Parameters assessed for rescue of lateral portion of Meckel’s cartilage (white boxes). Cells were scored as in B. n = 8–15 treated embryos over 4 experiments. (D) Experimental strategy involving injecting a morpholino targeting furina into mixed progeny of pmm2^m/m incross at 1-cell stage and genotyping 3 dpf. Confocal images of fli1a:EGFP-labeled cartilage structures show inhibiting furina improves chondrocyte morphology, organization, and differentiation in pmm2^m/m embryos. White boxes, central portion of Meckel’s cartilage; orange boxes, lateral regions evaluated for phenotypic rescue. Scale bars: 10 μm. Percentage values indicate the number of scored embryos exhibiting pictured phenotype. n = 8–15 treated embryos over 4 experiments. (E) Parameters assessed for rescue of central portion of Meckel’s cartilage (white boxes) following morpholino inhibition of furina. Cells were scored as in B. (F) Parameters assessed for rescue of lateral portion of Meckel’s cartilage (orange boxes) following morpholino inhibition of furina. Cells were scored as in B. Error bars show SEM, 2-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.0001, ****P < 0.0001.