Zebrafish sirt7 attenuates hypoxia signaling independent of its deacetylase activity.A, Western blot analysis of indicated protein levels in EPC cells transfected with empty vector, wildtype sirt7, or the enzyme-deficient mutants sirt7-S115A and sirt7-H191Y. B, luciferase activity of HRE-luciferase reporter in EPC cells transfected with empty vector, wildtype sirt7, or the enzyme-deficient mutants sirt7-S115A and sirt7-H191Y under normoxia (Nor) or hypoxia (Hyp). C, Western blot analysis of indicated protein levels in ZFL cells transfected with empty vector, wildtype sirt7, or the enzyme-deficient mutants sirt7-S115A and sirt7-H191Y. D, quantitative real-time PCR (qPCR) analysis of phd3 in ZFL cells transfected with empty vector, wildtype sirt7, or the enzyme-deficient mutants sirt7-S115A and sirt7-H191Y under normoxia (Nor) or hypoxia (Hyp). E, qPCR analysis of cited2 in ZFL cells transfected with empty vector, wildtype sirt7, or the enzyme-deficient mutants sirt7-S115A and sirt7-H191Y under normoxia (Nor) or hypoxia (Hyp). F, qPCR analysis of ldha in ZFL cells transfected with empty vector, wildtype sirt7, or the enzyme-deficient mutants sirt7-S115A and sirt7-H191Y under normoxia (Nor) or hypoxia (Hyp). p Values were calculated by two-way ANOVA analysis (B, D, E, and F); ∗∗p < 0.01, ∗∗∗p < 0.001 and ∗∗∗∗p < 0.0001; data based on one representative experiment performed in three biological replicates from at least three independent experiments (mean ± SD).
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