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Fig. 6 Ubiad1 and Coq2 Play Different Functions during Development(A) Se integrity is affected in Tg(kdrl:GFP)s843 embryos injected with ubiad1 morpholino (ubiad1 MO) but not in coq2 morphants (coq2 MO). Scale bar, 100 µm.(B) Oxidative stress levels in entire ubiad1 morphants (ubiad1 MO), coq2 morphants (coq2 MO), and relative control (ctrl MO).(C) CoQ10 levels detected by HPLC-MS are significantly lower in ubiad1 morphants (ubiad1 MO), coq2 morphants (coq2 MO), ubiad1 and coq2 double morphants (ubiad1+ coq2 MO) than control morphants (ctrl MO).(D) Embryos from bar heterozygote intercrosses were injected with coq2 mRNA and mitochondria-targeted CoQ10 analog called MitoQ (10 µM) or cherry mRNA and Decyl-TPP (10 µM) as respective controls. Phenotypes were scored at 72 hpf. bar phenotype is not caused by lack of mitochondria CoQ10.(E) Silencing of both CoQ10 biosynthetic enzymes (siUBIAD1 and siCOQ2) cause increase of total cellular ROS in ECs.(F) Levels of CoQ10 detected by HPLC-MS in total lipid extracts from HUAEC silenced for UBIAD1 (siUBIAD1), COQ2 (siCOQ2), or both (siUBIAD1+siCOQ2). UBIAD1 and COQ2 were silenced with 50 pmoles (black bars) or with 100 pmoles (gray bars). Levels of CoQ10 are significantly reduced in cells silenced for UBIAD1 or COQ2 expression.(G) Quantitative PCR analyses of UBIAD1 and COQ2 mRNA levels in HUAEC cells.(H) Quantification of loss of mitochondrial membrane potential (Δψm) in HUAEC silenced for UBIAD1 (siUBIAD1) or COQ2 (siCOQ2).(I) Quantification of lipid peroxidation by MDA adducts levels in HUAEC silenced for UBIAD1 (siUBIAD1) or COQ2(siCOQ2).All data are means ± SD. p < 0.01, p < 0.001.See also Figure S5.
Reprinted from Cell, 152(3), Mugoni, V., Postel, R., Catanzaro, V., De Luca, E., Turco, E., Digilio, G., Silengo, L., Murphy, M.P., Medana, C., Stainier, D.Y., Bakkers, J., and Santoro, M.M., Ubiad1 Is an Antioxidant Enzyme that Regulates eNOS Activity by CoQ10 Synthesis, 504-518, Copyright (2013) with permission from Elsevier. Full text @ Cell