Figure 3: Monitoring of H3K9ac-mintbody-GFP in response to changes in acetylation status in tobacco BY-2 cells.

(a,b) Time-lapse imaging of tobacco BY-2 cells expressing H3K9ac-mintbody-GFP (a) and GFP alone (b) in response to TSA treatment. The intensity of GFP fluorescence is shown by a heat map. The same cells were observed at the indicated time point. Bars, 100 μm. (c,d) Quantitative analysis of GFP levels in tobacco BY-2 cells expressing H3K9ac-mintbody-GFP (c) and GFP alone (d) treated with TSA for 3 h. The ratio of nuclear/cytoplasmic intensity of GFP is shown. The values represent means ± S.D. (n = 13–16, t-test; **p < 0.01, N.S., not significant). (e,f) Time-lapse imaging of tobacco BY-2 cells expressing H3K9ac-mintbody-GFP (e) and GFP alone (f) in response to C646 treatment. The intensity of GFP fluorescence is shown by a heat map. The same cells were observed at the indicated time point. Bars, 100 μm. (g,h) Quantitative analysis of GFP levels in tobacco BY-2 cells expressing H3K9ac-mintbody-GFP (g) and GFP alone (h) treated with C646 for 3 h. The ratio of nuclear/cytoplasmic intensity of GFP is shown. The values represent means ± S.D. (n = 14–18, N.S., not significant by t-test). (i) Chemical structures of synthesized HDAC inhibitors Ky-2 and Ky-14. (j) Quantitative analysis of GFP levels in tobacco BY-2 cells expressing H3K9ac-mintbody-GFP treated with Ky-2 and Ky-14 for 3 h. The ratio of nuclear/cytoplasmic intensity of GFP is shown. The values represent means ± S.D. (n = 34–38, t-test; *p < 0.05, **p < 0.01).