Figure 3

Garcinol induces DNA repair pathways and alters p53 acetylation. (A) Immunostaining of MCF7 cells for the DNA damage marker γH2A.X following treatment with curcumin, garcinol or vehicle for 24 hours. (B) Western blots of acid extracted histones prepared from MCF7 cells following treatment for 24 hours with HAT inhibitors at the indicated concentrations. Specific antibodies were used to reveal γH2A.X and H3K56Ac. Immunodetection of histone H3 and Coomassie staining of extracted histones are shown as loading controls. (C) Western blots showing the levels of hMOF, TIP60 and SIRT1 proteins in whole cell extracts of MCF7 cells following curcumin or garcinol treatment or control (DMSO). (D) Western blots detecting p53 expression levels and selected p53 acetylation PTMs (K373/382Ac or K120Ac) following garcinol treatment of MCF7 cells for 24 hours. (E) Immunostaining of MCF7 cells as treated in (D) showing the detection level and subcellular distribution of p53 and TIP60 proteins in MCF7 cells. Scalebar is 10 μm. (F) Higher magnification of boxed insets in (E) showing localisation of p53K120Ac and p53K373/382Ac proteins to the cytoplasm and nucleus, respectively.