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Fig. 4. The levels of H3(K9) dimethylation (17 kDa), H3(K9) acetylation (17 kDa) and H3(K4) dimethylation (17 kDa) analysed by western blots in (A) the human HT29 colon cancer cell line, and (B) normal colon cells FHC. In order to compare the effects of HDACi on normal and tumour cells, the following treatments were tested: sodium butyrate (HT29/NaBt; FHC/NaBt), TNF{alpha} (HT29/TNF{alpha}; FHC/TNF{alpha}) and a combination of NaBt with TNF{alpha} (HT29/NaBt+TNF; FHC/NaBt+TNF). In both cell types, in comparison with the control, there was a substantial increase in H3(K9) and H3(K4) dimethylation and H3(K9) acetylation after treatments involving NaBt. Apoptotic cleavage of lamin B (65 kDa) into a 45 kDa fragment was enhanced after the combined treatment with NaBt and TNF{alpha} (see A and B). Protein levels for all treatments were normalized to the selected band (arrow) of total nuclear protein levels. (C) Comparison of the effects of TSA and NaBt on H3(K9) dimethylation (17 kDa), H3(K9) acetylation (17 kDa) and H3(K4) dimethylation (17 kDa) of HT29 cells. (D) Immunocytochemical analyses of H3(K9) dimethylation, H3(K9) acetylation and H3(K4) dimethylation patterns in HT29 cells treated with TSA and NaBt. TO-PRO-3 was used as a counterstain. The images represent the histone modification patterns induced by HDACi in the majority of the cell population. White and yellow arrows indicate the nuclear periphery and the nucleoli, respectively. Bars, 1 µm.