Skip to main content
. 2013 Oct 1;5(11):1662–1683. doi: 10.1002/emmm.201202055

Figure 8. TIP60 regulates p53-mediated activation of BAX or NOS3 gene in H9c2, H9c2 p53−/− and H9c2 p53-Lys118 (Mut) cells.

Figure 8

  1. TIP60 protein expression in H9c2 cells under control, SD, and SD + Oxy conditions. SD induces TIP60 expression, which is inhibited by oxygenation.
  2. TIP60 expression in MI hearts is inhibited by oxygenation.
  3. Co-immunoprecipitation of p53 with TIP60 shows that TIP60 binds to p53 only in MI hearts and this interaction is abolished upon oxygenation. Input and SD-treated H9c2 cells with TIP60 gene-silencing were used as controls.
  4. Effect of TIP60 on the survival of SD H9c2 cells. Silencing of TIP60 using TIP60 siRNA in H9c2, H9c2 p53−/− and H9c2 p53-Lys118(Mut) cells increases the survival of SD cells. Addition of TIP60 using cDNA results decreases the survival of SD cells. Data represent mean ± SD of four independent measurements.
  5. Effect of TIP60 on the binding of p53 to BAX-RE and NOS3-RE was analysed using ChIP in H9c2 cells. Results show that TIP60 gene-silencing results in binding of p53 to NOS3-RE in both SD and SD + Oxy cells. Similarly, exogenous addition of TIP60 cDNA results in binding of p53 to BAX-RE in both SD and SD + Oxy cells.
  6. Effect of TIP60 siRNA and TIP60 cDNA addition on p53-Lys118 acetylation, NOS3 and BAX protein expression was determined using Western blot. Data show that TIP60 siRNA abolishes p53-Lys118 acetylation, increases the expression of NOS3 protein and decreases BAX expression in H9c2 cells. Similarly, TIP60 cDNA increases p53-Lys118 acetylation, decreases the expression of NOS3 protein and increased BAX expression in H9c2 cells.
  7. Expression of p53 downstream genes was determined in the SD and SD + Oxy group of H9c2 cells in presence and absence of TIP60 siRNA and TIP60 cDNA. Data show that TIP60 cDNA increases the expression of these genes and TIP60 siRNA decreases their expression in both SD and SD + Oxy groups.