Fig. 5
NHR-14 cooperates with CEP-1/p53 to regulate egl-1 and ced-13 expression. A Relative fold induction of egl-1 mRNA in N2, nhr-14(tm1473), gld-1(op236) and gld-1(op236); nhr-14(tm1473) after 24 h of gamma irradiation (120 Gy). egl-1 induction was averaged from three independent RT-qPCR analyses. B Relative fold induction of ced-13 mRNA in N2, nhr-14(tm1473), gld-1(op236) and gld-1(op236); nhr-14(tm1473) following 24 h of gamma irradiation (120 Gy). egl-onefold change was averaged from three independent RT-qPCR analyses. C Western blotting analysis of CEP-1 levels in N2 and nhr-14(tm1473) animals. D NHR-14 interacts with CEP-1/p53 in mammalian cells. Flag-NHR-14 and Myc-CEP-1/p53 were co-expressed in HEK293 cells and then immunoprecipitated (IP) using Flag antibodies. The immunoprecipitated proteins were detected by immuno-blotting (IB) with Myc antibodies. E. NHR-14 and CEP-1/p53 directly interact in vitro. The full-length NHR-14 protein was in vitro translated and labeled with [.35S] methionine and incubated with GST or GST-CEP-1/p53 fusion proteins, which were immobilized on glutathione sepharose beads, for 2 h. After extensive washes, the bound proteins were resolved by SDS-PAGE and detected by autoradiography