FIGURE 5.

TIP60 interacts with MRTF-A to activate iNOS transcription. (A) RAW264 cells were exposed to hypoxia-reoxygenation. ChIP assays were performed with anti-TIP60 or IgG. (B) HEK293 cells were transfected with FLAG-MRTF-A and/or Myc-tagged TIP60. Immunoprecipitation was performed with anti-FLAG. (C) Whole cell lysates from RAW cells were immunoprecipitated with anti-MRTF-A or IgG. (D) RAW264 cells were exposed to hypoxia-reoxygenation. Re-ChIP assays were performed with indicated antibodies. (E) An iNOS promoter construct was transfected into HEK293 cells with MRTF-A and/or TIP60. Luciferase activities were normalized by protein concentration and GFP fluorescence. (F,G) RAW264 cells were treated with MG149 and/or hypoxia-reoxygenation. Expression levels of iNOS were examined by qPCR and Western. (H–K) RAW264 cells were transfected with siRNA targeting TIP60 or scrambled siRNA (SCR) followed by exposure to hypoxia-reoxygenation. Expression levels of iNOS were examined by qPCR and Western. ChIP assays were performed with anti-acetyl H4K16 and anti-MRTF-A. N = 3 for all the experiments. Data represent averages of three independent experiments and error bars represent SEM.