Figure 5

Zebrafish sirt7 binds to hif-1α and hif-2α.A, coimmunoprecipitation analysis of Myc-sirt7 with Flag-hif-1αa, or Flag-hif-1αb, or Flag-hif-2αa, or Flag-hif-2αb. HEK293T cells were transfected with Myc-sirt7, together with Flag empty vector, or Flag-hif-1αa, or Flag-hif-1αb, or Flag-hif-2αa, or Flag-hif-2αb. Anti-Flag-conjugated agarose beads were used for coimmunoprecipitation, and the indicated antibodies were used for detection. B, coimmunoprecipitation analysis of Myc-sirt7 with Flag-hif-1αa under normoxia (Nor) or hypoxia (Hyp). HEK293T cells were transfected with Myc-sirt7, together with Flag empty vector or Flag-hif-1αa, and cultured under normoxia (Nor) or hypoxia (Hyp) for 4 h. Anti-Flag-conjugated agarose beads were used for coimmunoprecipitation, and the indicated antibodies were used for detection. The Flag-hif-1αa protein under hypoxia was adjusted to be similar to that under normoxia. The immunoprecipitated Myc-sirt7 protein by Flag-hif-1αa in IP (∗) over the Myc-sirt7 protein in TCL (#) was determined (∗/#). C, coimmunoprecipitation analysis of Myc-sirt7 with Flag-hif-1αb under normoxia (Nor) or hypoxia (Hyp). HEK293T cells were transfected with Myc-sirt7, together with Flag empty vector or Flag-hif-1αb, and cultured under normoxia (Nor) or hypoxia (Hyp) for 4 h. Anti-Flag-conjugated agarose beads were used for coimmunoprecipitation, and the indicated antibodies were used for detection. The Flag-hif-1αb protein under hypoxia was adjusted to be similar to that under normoxia. The immunoprecipitated Myc-sirt7 protein by Flag-hif-1αb in IP (∗) over the Myc-sirt7 protein in TCL (#) was determined (∗/#). D, coimmunoprecipitation analysis of Myc-sirt7 with Flag-hif-2αa under normoxia (Nor) or hypoxia (Hyp). HEK293T cells were transfected with Myc-sirt7, together with Flag empty vector or Flag-hif-2αa, and cultured under normoxia (Nor) or hypoxia (Hyp) for 4 h. Anti-Flag-conjugated agarose beads were used for coimmunoprecipitation, and the indicated antibodies were used for detection. The Flag-hif-2αa protein under hypoxia was adjusted to be similar to that under normoxia. The immunoprecipitated Myc-sirt7 protein by Flag-hif-2αa in IP (∗) over the Myc-sirt7 protein in TCL (#) was determined (∗/#). E, coimmunoprecipitation analysis of Myc-sirt7 with Flag-hif-2αb under normoxia (Nor) or hypoxia (Hyp). HEK293T cells were transfected with Myc-sirt7, together with Flag empty vector or Flag-hif-2αb, and cultured under normoxia (Nor) or hypoxia (Hyp) for 4 h. Anti-Flag-conjugated agarose beads were used for coimmunoprecipitation, and the indicated antibodies were used for detection. The Flag-hif-2αb protein under hypoxia was adjusted to be similar to that under normoxia. The immunoprecipitated Myc-sirt7 protein by Flag-hif-2αb in IP (∗) over the Myc-sirt7 protein in TCL (#) was determined (∗/#). IP, immunoprecipitation; TCL, total cell lysates.