Fig. 3: Transcriptional changes and anti-leukemia effects induced by catalytic histone acetylation.

Before analysis, cells were treated with catalyst 1 (acetylated) or 2 (control) (10 µM) and acetyl donor 4 (1 mM) for 4 h. a, b Correlation between catalytic histone acetylation and gene transcription. After acetylation for 4 h, THP-1 cells were analyzed by RNA-seq (n = 2). Red and blue dots represent significantly upregulated and downregulated genes, respectively (P < 0.05 in Student’s two-sided t test, log₂FC > 1 or < 1). Gray dots represent other genes (NS). a Scatter plot of the FPKM + 1 value of genes. b Scatter plot of the FPKM + 1 value of MLL-fusion target genes (Human Gene Set: KUMAR_TARGETS_OF_MLL_AF9_FUSION). c, d Correlation between catalyst-promoted acetylation and cell cycles or apoptosis. After acetylation for 4 h, the medium was replaced with fresh medium. After 18 h, cells with different cell cycles (G0/G1, S, or G2/M) were classified by flow cytometry for DNA content (c). After 48 h, cells with early apoptosis (early apop.: high Annexin V and low 7-AAD), late apoptosis (late apop.: high Annexin V and high 7-AAD), or dead (low Annexin V and high 7-AAD) were classified by flow cytometry for Annexin V and 7-AAD staining (d). The bar graph shows the mean of three independent experiments. e, f Correlation between catalytic histone acetylation and viability of cell lines. After acetylation for 4 h, the medium was replaced with fresh medium. The numbers of trypan blue-negative cells at 0–6 days are shown, normalized to the cells treated with DMSO in (e), or the cells with 2 + 4 in (f). Data are shown as mean ± standard deviation of four independent experiments (*P < 0.05, one-way ANOVA with Tukey’s post hoc test to DMSO-treated cells in (e), or 4-treated cells in (f). NS: not significant). g, h Correlation between catalytic histone acetylation and the tumorigenic potential in mice. After acetylation for 4 h, Akaluc-expressing 3 × 10⁶ MOLM-13 cells were intravenously injected into NOG mice (n = 5 for control, 6 for acetylated). g Images of bioluminescence signals in recipient mice at 7 and 14 days post-transplantation. h A Kaplan-Meier survival. Statistical significance was calculated with log-rank tests.