Fig 2: Effect of compound 5 on the acetylation status of Histone H3 and α-tubulin. MDA-MB-231 cells were incubated with the indicated concentrations of compound 5 for 24 hours and the expression level of Histone H3, Ac-Histone H3, α-tubulin, Ac-α-tubulin, HDAC1, HDAC6, and GAPDH were analyzed using Western blotting. DMSO (D) was used as a negative control. GAPDH = glyceraldehyde 3-phosphate dehydrogenase, HDAC = Histone deacetylase.

Fig 3: LMK235 decreases CaMKIIα overexpression‐induced cell cycle genes and class I/IIa HDACs. (A‐D) A10 cells were transfected with pCMV‐SPORT6‐CaMKIIα or pCMV‐SPORT6 empty vector and treated with LMK235 (1 μmol/L) for 24 h. Transcript levels of CaMKIIα, cyclin D1 and class I/IIa HDACs were determined by qRT‐PCR. Target genes were normalized to 18S rRNA. Data are expressed as mean ± SE of three independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 vs empty vector; ## P < 0.01 and ### P < 0.001 vs CaMKIIα transfection group; NS indicates not significant. (E and F) Western blot analyses of cyclin D1, HDAC1, HDAC2, HDAC4 and HDAC5. GAPDH was used as a loading control. The amount of protein expression was quantified by densitometry. *P < 0.05 vs empty vector; # P < 0.05, ## P < 0.01 and ### P < 0.001 vs CaMKIIα transfection group

Fig 4: Analysis of gene knockout effects in HCT116 cells and gene expression changes in melanoma associated with corin treatment. a Knockout of LSD1 in HCT116 cells decreased the rate of cancer cell proliferation by ~40% (n = 4). b, c Knockout of LSD1 enhanced the potency of MS-275 but did not affect the potency of corin in HCT116 cells as determined by [3H]thymidine incorporation after 48 h treatment (n = 4). d, e Venn diagrams depicting increases in global and tumor suppressor gene transcription (fold change = 2s, p < 0.05), respectively, upon 2.5 µM MS-275 or corin treatment (n = 3). f, g qRT-PCR validation of gene expression changes identified by microarray. WM983B melanoma cells were treated with inhibitor (2.5 µM) for 24 h prior to RNA isolation and analysis (n = 3). h ChIP-PCR localized CoREST complex components HDAC1 and LSD1 to the promoter region of CHOP (n = 3). i ChIP-PCR localized HDAC1, but not LSD1, to the promoter region of SYN1 (n = 3). Data (mean ± SEM) are representative of at least three independent experiments (unpaired t test, *p < 0.05, ***p < 0.001)

Fig 5: Model. a In healthy lung fibroblasts, C1D-bound MIRLET7D-ncRNA duplexes attract active HDAC1/2 and the MiCEE complex to specific loci. Active HDAC1/2 deacetylate H3K27, whereas EZH2 mediates trimethylation of H3K27, thereby leading to heterochromatin formation and transcriptional silencing. b In IPF lung fibroblasts, reduced MIRLET7D and active EP300, which inactivates HDAC1/2 and interacts with C1D, disrupt the MiCEE complex interfering with heterochromatin formation and transcriptional silencing at specific loci