Figure 1
Omomyc and Myc-dependent H4R3 symmetric di-methylation.
(A) Western blot analysis of U87MG/FO cell extracts along a time course between 3 and 24 hrs after doxycycline treatment. Picture shows the global changes of several histone modifications upon Omomyc expression. H3K9 histone acetylation decreased, as expected. H3K27 tri-methylation slightly decreased, as well. On the contrary, a peak of K4 tri-methylation on histone H3 was observed after 8 hrs of doxycycline treatment. The increase in H4R3 symmetric di-methylation followed the kinetic of expression of Flag-Omomyc. (B) Western blot analysis of BT168/FO and BT275/FO cell extracts along a time course between 3 and 48 hrs after doxycycline treatment. H4R3me2s increased in parallel with Flag-Omomyc. (C) Western blot analysis of nuclear extracts from HEK293T cells transfected with Flag-Omomyc and Flag-Myc encoding vectors. H4R3me2s was induced upon both Omomyc and Myc expression. The empty vector was used as control. (D) Double immunofluorescence. HEK293T cells transfected with either Flag-Omomyc or Flag-Myc enconding vectors were stained simultaneously with anti-H4R3me2s and anti-Flag antibodies. HEK293T cells expressing either Flag-Omomyc or Flag-Myc were also positive for H4R3me2s. These results are representative of at least three independent experiments.
