Fig. 5: CR recruits HDAC3 to promote aberrant myeloid differentiation.

a The interaction between CR fusions and HDAC3 in human CD34⁺ cells expressing CR. * indicate degraded flag-CRL proteins. M, markers. b Venn diagram displaying the shared peaks of CR fusions and HDAC3 in CD34⁺ cells expressing CRL (top panel) or CRS (bottom panel). c The heatmaps showing the signal intensities at DNA loci associated with common peaks of CR-HDAC3 in CD34-CR cells, both untreated and treated with RGFP966 or Tucidinostat. d–e The colony-forming of CD34⁺ cells expressing Vec, CRL or CRS following silencing of HDAC3 shRNAs (d) or treatment with HDAC inhibitors (e). n = 3 independent donors in n = 3 independent experiments. f–g Percentage of CD66⁺ cells assessed by flow cytometry in CR-expressing CD34⁺ cells after silencing of HDAC3 shRNAs (f, n = 3 independent donors in n = 3 independent experiments) or treatment with HDAC inhibitors (g, n = 3 independent donors in n = 4 independent experiments). h–i HDAC inhibitors lead to significantly decreased CR binding at PU.1 promoter in CD34-CRL (h) and CD34-CRS cells (i). j RGFP966 or tucidinostat reversed the trans-repression of PU.1. n = 4 biological replicates. k The protein levels of PU.1 and acetylated histone H3 (Ac-H3) in CD34-Vec, CD34-CRL and CD34-CRS cells following silencing of HDAC3 shRNAs. l Silencing of PU.1 reduced the apoptosis induced by HDAC inhibitors in HL60-CR cells. n = 3 biological replicates. The data are shown as the mean ± SD, statistical analysis was performed by two-tailed one-way ANOVA with Dunnett’s (d–g, l) or Tukey’s (j) multiple comparisons test. The images are representative of three independent experiments from three healthy donors (a, k). See also Supplementary Figs. 6–8. Source data are provided as a Source Data file.