Fig. 6.

Breast cancer cells and tumors display high basal histone glycation and sensitivity to MGO. a Indicated breast cancer cell lines (and 293T as a reference) were untreated or treated with 0.25, 0.5 or 1 mM MGO for 12 h, after which histones were extracted by high salt and analyzed by western blot on the same SDS-PAGE with the indicated antibodies. b SKBR3 breast cancer cell line was pre-treated for 2 h with 10 mM carnosine, after which increasing amounts of MGO were added to the media. Histones were extracted as described above and samples were analyzed by western blot with anti-MGO. c 293T cells were pre-treated with histone acetyl-transferases (curcumin, 50 μM) or deacetylases (SAHA, 5 μM) inhibitors for 2 h after which increasing amounts of MGO were added to the media. Histones were extracted as described above and samples were analyzed by western blot with the indicated antibodies. d Indicated breast cancer xenografts tumors were analyzed for histone glycation, by extracting the histones with high salt and analyzing them by western blot with the indicated antibodies. Corresponding cytosolic glycation analysis and DJ-1 expression can be found in Supplementary Figure 11. e Non-tumor (NT) and tumor (T) samples were obtained from five different breast cancer patients (P1-P5) and both soluble and histone fractions were extracted and analyzed as described above. Corresponding DJ-1 expression levels are found in Supplementary Figure 11