American Society of Hematology

Fig. 3.

$Fig. 3. Effect of HDAC inhibitors on K562 cells and histone hyperacetylation in ILT-Mat and K562 cells in response to butyrate. / (A) K562 cells were cultured for 48 hours without (open column) or with sodium butyrate (shaded column) or TSA (closed column) at the indicated concentrations (mmol/L or nmol/L, respectively). Cell viability was assessed by trypan blue dye exclusion assay. The results, 7.3 ± 0.8(0), 10.6 ± 3.8 (0.5), 16.6 ± 3.6(1), 19.4 ± 4.4(2), 8.4 ± 1.0(200), and 16.2 ± 0.6%(500) of total cells, are averages of triplicate samples from 3 experiments ± SD. (B) Histone acetylation in ILT-Mat and K562 cells after butyrate treatment was evaluated by fractionating histones on acid/urea/acrylamide gels. Cells (5 × 106) were exposed to sodium butyrate at the indicated concentration (mmol/L) for 18 hours. Histones were extracted and loaded onto SDS-PAGE gels that were fixed and stained in 0.25% Coomassie blue/10% acetic acid/40% methanol. The lowest band in each lane is unacetylated H4, and the numbers to the right indicate the number of acetylated lysines in H4.$

Effect of HDAC inhibitors on K562 cells and histone hyperacetylation in ILT-Mat and K562 cells in response to butyrate.

(A) K562 cells were cultured for 48 hours without (open column) or with sodium butyrate (shaded column) or TSA (closed column) at the indicated concentrations (mmol/L or nmol/L, respectively). Cell viability was assessed by trypan blue dye exclusion assay. The results, 7.3 ± 0.8(0), 10.6 ± 3.8 (0.5), 16.6 ± 3.6(1), 19.4 ± 4.4(2), 8.4 ± 1.0(200), and 16.2 ± 0.6%(500) of total cells, are averages of triplicate samples from 3 experiments ± SD. (B) Histone acetylation in ILT-Mat and K562 cells after butyrate treatment was evaluated by fractionating histones on acid/urea/acrylamide gels. Cells (5 × 10⁶) were exposed to sodium butyrate at the indicated concentration (mmol/L) for 18 hours. Histones were extracted and loaded onto SDS-PAGE gels that were fixed and stained in 0.25% Coomassie blue/10% acetic acid/40% methanol. The lowest band in each lane is unacetylated H4, and the numbers to the right indicate the number of acetylated lysines in H4.

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