Fig 2: Sodium butyrate inhibits the expression of type I HDAC in THP-1 cells infected with M. bovis. THP-1 cells were treated with different concentration of NaB (0, 0.5, and 1.0 mM) for 24 h followed by infection with M. bovis (MOI 10:1). (A) The expression of HDAC1, HDAC2, and HDAC3 were determined by western bolt assay. The relative band density of (B) HDAC1, (C) HDAC2, and (D) HDAC3 were calculated by using GAPDH as internal control. Data are representative of three independent experiments and values are expressed as mean ± SD. *P < 0.05; **P < 0.01.

Fig 3: Chidamide regulates the expression of lncRNAs and inhibits the oncogenic MAPK signaling pathway in AML cells. (A) Cluster analysis of lncRNAs differentially expressed in Chidamide-treated group and control group. Whether the cells were treated with Chidamide at 1,000 nM for 48 h. (B) 1,195 lncRNAs were co-upregulated and 780 lncRNAs were co-downregulated were observed. (C) Expression of VPS9D1-AS1 in the Chidamide-treated group and the control group analyzed by qRT-PCR. Each bar presents mean ± SD from three independent experiments * p < 0.05, ** p < 0.01. (D) Overall survival was analyzed and compared by Kaplan-Meier. Patients with high (lncRNA VPS9D1-AS1 high, n = 53) vs. low (lncRNA VPS9D1-AS1 low, n = 53). (E) Polymerase chain reaction analysis of lncRNA VPS9D1-AS1 expression in patients with de novo AML (n = 21) vs. health control people (n = 27). * p < 0.05, ** p < 0.01, *** p < 0.001. (F) Pathway enrichment analysis showing genes involved in MAPK signaling pathway were significantly enriched. (G) Cluster analysis was performed on genes related to the oncogenic MAPK signal. (H) GSEA showed that oncogenic MAPK signaling was down-regulated in Chidamide-treated AML cells. (I) Protein network analysis revealed top 10 of the enriched oncogenic MAPK signal proteins were closely connected to HDAC1/2/3/8/10/11.

Fig 4: Repression of PTEN by MTA2 is dependent on Snail.a The luciferase reporter assay in HEK-293T cells co-transfected with the wild-type (WT) or mutant (Mut) PTEN promoter luciferase reporter and the vector or MTA2 constructs. Schematic of the sequence of the putative consensus MTA2-binding element in the human PTEN promoter region and the substitution mutations introduced into this binding element sequence are shown. The luciferase reporter activity results were depicted as a bar graph with mean ± S.D. n = 3. *P < 0.05. b qChIP assays in MIA Paca-2 cells or PANC-1 cells were performed for the presence of MTA2 at the PTEN promoter with or without MTA2 depletion. Diagram of the PTEN promoter region with four amplicons used for qPCR analysis. Error bars represent mean ± S.D. n = 3. **P < 0.01. c Endogenous Snail or HDAC1 level was measured in MIA Paca-2 cells or PANC-1 cells following transduction with Snail shRNAs (shSnail) or HDAC1 shRNAs (shHDAC1) and shSCR (negative control). GAPDH served as an internal control for each group. Values are mean ± S.D. n = 3. **P < 0.01. d MIA Paca-2 cells were infected with lentiviruses carrying the indicated shRNAs. A qChIP assay was performed using specific antibodies against MTA2, Snail, HDAC1, H3Ac, or H3 to detect their binding onto the PTEN promoter. Error bars represent mean ± S.D. for three independent experiments. *P < 0.05; **P < 0.01. e qRT-PCR analyses were used to measure the expression of PTEN in MIA Paca-2 or PANC-1 cells transfected with shSCR or shSnail. f The expression of PTEN was measured by qRT-PCR in MIA Paca-2 or PANC-1 cells co-transfected with shSnail and the expression construct for MTA2. g ChIP and Re-ChIP experiments in MIA Paca-2 cells with the antibodies against Snail and MTA2 or with isotypic IgG as negative controls

Fig 5: RACK1 maintains co-repressor HDAC1 protein level in hepatocytes. (A) BNL CL.2 cells were transfected with the indicated siRNA. Whole-cell lysates were harvested 48 h later and subjected to IB analysis with indicated antibodies. ns, non-specific band. (B-D) Livers of 12-week-old Rack1?hep mice and littermate controls were subjected to IB (B), IHC (C, scale bar, 50 µm), and quantitative RT-PCR (D) analysis to examine the expression of HDAC1. (E, F) ChIP assays of HDAC1 recruitment (E) and histone H3K9 and H3K27 acetylation (F) at the Cxcl1 and S100a9 promoters in livers of 12-week-old Rack1?hep mice and littermate controls. Error bars indicate mean ± SD. The n-value information is shown in the corresponding panel. *p < 0.05; **p < 0.01; ***p < 0.001; NS, not significant.