Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: EZH2 and HDAC1/2 expression in PTCL.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: Representative immunohistochemical features of HDAC1 (left), HDAC2 (middle), and EZH2 (right) in PTCL-NOS, ALCL, NK/T and AITL. All images were captured at ×200 and ×400 magnifications. HDAC, histone deacetylase; EZH2, enhancer of zeste homolog 2; PTCL-NOS, peripheral T cell lymphoma not otherwise specified; ALCL, anaplastic large cell lymphoma; NK/T, natural killer/T-cell; AITL, angioimmunoblastic T-cell lymphoma.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques: Immunohistochemical staining, Histone Deacetylase Assay

Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: Correlations between EZH2 and HDAC1/2 in four subtypes of PTCL.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques:

Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: Correlations between EZH2/HDAC1/2 expression and the clinicopathological characteristics in PTCL.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: Correlations between the EZH2/HDAC1/2 expression and the clinicopathological characteristics in PTCL-NOS.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: Correlations between the EZH2/HDAC1/2 expression and the clinicopathological characteristics in NK/TCL.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: OS rates based upon the expression levels of EZH2 and HDAC1/2 in PTCL. (A) Significant trend towards poorer OS rates for patients with high EZH2 expression (P=0.012). (C) High expression of HDAC2 was correlated with a poorer OS rate compared with low expression of HDAC2 (P<0.001), which was not observed in the HDAC1 group [(B); P=0.353]. OS, overall survival; HDAC, histone deacetylase; EZH2, enhancer of zeste homolog 2.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques: Expressing, Histone Deacetylase Assay

Journal: Oncology Letters

Article Title: Clinical significance of enhancer of zeste homolog 2 and histone deacetylases 1 and 2 expression in peripheral T-cell lymphoma

doi: 10.3892/ol.2019.10410

Figure Lengend Snippet: OS based upon the levels of EZH2 and HDAC1/2 in PTCL-NOS. (A) Significant trend towards poorer OS rates for patients with high EZH2 (P=0.002). (C) High expression of HDAC2 was correlated with a poorer OS rate, compared with low expression of HDAC2 (P<0.001), which was not observed in the HDAC1 group [(B); P=0.339]. OS, overall survival; HDAC, histone deacetylase; EZH2, enhancer of zeste homolog 2; PTCL-NOS, peripheral T cell lymphoma not otherwise specified.

Article Snippet: Following rinsing with phosphate-buffered saline, the slides were incubated with polyclonal rabbit anti-HDAC1 (catalog no. BS6485; 1:100 dilution; Bioworld Technology, Inc., St. Louis Park, MN, USA), polyclonal rabbit anti-HDAC2 (catalog no. 12922-3-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and polyclonal rabbit anti-EZH2 (catalog no. BS90776; 1:50 dilution; Bioworld Technology, Inc.) primary antibodies overnight at 4°C.

Techniques: Expressing, Histone Deacetylase Assay

Journal: Clinical Cancer Research

Article Title: Zinc Finger MYND-Type Containing 8 (ZMYND8) Is Epigenetically Regulated in Mutant Isocitrate Dehydrogenase 1 (IDH1) Glioma to Promote Radioresistance

doi: 10.1158/1078-0432.ccr-22-1896

Figure Lengend Snippet: Figure 4. ZMYND8 knockout GCCs display reduced viability to irradiation, which is further enhanced when combined with BRD4 and HDAC inhibition. A, Experimental model in which ZMYND8 lentiviral particles were generated to knockout (KO) ZMYND8 expression in human and mouse GCCs mediated by CRISPR-Cas9-sgRNAs. ZMYND8 KO GCCs were selected based on resistance to 10 mg/mL puromycin for 1 week in mouse mIDH1 GCCs and 2 weeks for human mIDH1 GCCs. Representative Western blot quantification of ZMYND8 KO for (B) SF10602, (C) MGG119, and (D) NPAI mouse NS. Cellular viability of ZMYND8 WT, shown by a black line vs. ZMYND8 KO, represented by the red line, was assessed 72 hours after irradiation (IR) exposure using CellTiter-Glo assay in the (E) SF10602, (F) MGG119, and (G) NPAI. Results are expressed in relative luminescence units (RLU) to control nonirradiated (0 Gy) cells. H, Working model of ZMYND8-interacting partner bromodomain-containing protein (BRD4), where ZMYND8 is recruited to enhancer regions marked by H3K4me1 and contributes to the regulation of cancer cell survival and proliferation-associated genes. BRD4 inhibitors, JQ1 and I-BET-762, can disrupt this interaction. I, Working model of ZMYND8-interacting partner HDAC1/2 (histone deacetylase 1/2); a component of the NuRD complex, where ZMYND8 binds H3K14ac residues present at damaged chromatin regions and recruits HDAC along with other NuRD subunits: MDB2, CHD4, and GATAD2A. Panobinostat inhibits HDAC1/2 and prevents histone deacetylation mediated by HDAC1/2, which is required for transcriptional repression at regions of DNA damage. J, Representative bar graph of cellular viability measured in RLU, which shows the effect of BRD4 (JQ1, I-BET-762) or HDAC (panobinostat) inhibition alone (IR) or in combination with irradiation (þIR) in mouse NPAI ZMYND8 WT in blue vs. NPAI ZMYND8 KO in red. Errors bars represent SEM from independent biological replicates (n ¼ 3). ns, not significant; , P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001; unpaired t test.

Article Snippet: After blocking, membranes were incubated with primary anti-phospho gH2AX (1:1,000; Cell Signaling Technologies, cat. #9718S), anti-gH2AX (1:1,000; Cell Signaling Technologies, cat. #2595S), primary antibody ZMYND8 (1:2,000; Bethyl Laboratories, cat. #A302-089A), primary antibody TIMELESS (1:1,000; Bethyl Laboratories, cat. #A300-960A), primary antibody proliferating cell nuclear antigen (PCNA) (1:1,000; Cell Signaling Technologies, cat. #2586T), primary antibody TREX1 (1:1,000; Abcam, cat. #ab185228), primary antibody BRD4 (1:1,000; Cell Signaling Technology, cat. #83375B), primary antibody HDAC1 (1:1,000; Cell Signaling Technology, cat. #2062S), primary antibody HDAC2 (1:1,000; Cell Signaling Technology, cat. # 57156S), Vinculin (1:3,000; Thermo Fisher Scientific, cat. #7,000,062), or b-tubulin antibodies (1:4,000; Sigma-Aldrich, cat. #A1978) overnight at 4 C. The next day, blots were washed with TBS-0.1% tween-20 and incubated with secondary (1:4,000) antibodies [Dako, Agilent Technologies, goat anti-rabbit 1:4,000 (cat. # P0448), rabbit anti-mouse 1:4,000 (cat. # P0260)] for 1 hour at RT.

Techniques: Knock-Out, Irradiation, Inhibition, Generated, Expressing, CRISPR, Western Blot, Glo Assay, Control, Histone Deacetylase Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: CBX2 suppresses interferon signaling to diminish tumor immunogenicity via a noncanonical corepressor complex.

doi: 10.1073/pnas.2417529122

Figure Lengend Snippet: Fig. 5. RACK1 works as the essential scaffold for the CBX2–HDAC1 complex to perform deacetylation. (A) Co-IP and immunoblot analysis of the interaction of HDAC1 and RACK1 in extracts of B16 OVA cells. (B) The expression of HDAC1, CBX2, and RACK1 in control and Cbx2-KD B16 OVA tumor cells was detected by WB. β-actin was used as the loading control. (C) Co-IP and immunoblot analysis of the interaction of the HDAC1 and RACK1 in extracts of Cbx2-KD or control B16 OVA cells. (D) Co-IP and immunoblot analysis of the interaction of HDAC1 and CBX2 in extracts of B16 OVA cells. (E) Flag-CBX2 expressed B16 OVA cells were stained with anti-Flag (green) and anti-HDAC1 (red) antibodies to detect CBX2 and HDAC1 via confocal microscopy. (Scale bars, 10 μm.) (F) Co-IP and immunoblot analysis of the interaction of the HDAC1 and CBX2 in extracts of Rack1-KO or control B16 OVA cells. (G) The distribution of HDAC1, CBX2, and RACK1 in control and Cbx2-KD B16 OVA tumor cells was detected by WB. β-actin was used as the whole cell loading control. GAPDH was used as cytosol protein control, and Histone H3 was used as chromatin-binding protein control. (H) ChIP-qPCR analysis of HDAC1 occupancy at genomic loci of Cxcl9 and Cxcl10 in Cbx2-KD and control B16 OVA cells mediated by shRNA. (I) Schematic model for the role of CBX2 in suppressing tumor immunogenicity. **P < 0.01, ***P < 0.001, in one-way ANOVA with Bonferroni’s posttest (H).

Article Snippet: The following antibodies were used for western blot analyses: β- actin (Santa Cruz, catalog sc8432), Flag (Sigma, catalog A8592), HA (Roche, 12013819001), CBX2 (CST, catalog 18687), RACK1 (CST, catalog 5432), HDAC1 (CST, catalog 34589), EZH2 (CST, catalog 5246), GAPDH (Santa Cruz, catalog sc32233), H3K27ac (CST, catalog 8173), and H3K27me3 (CST, catalog 9733).

Techniques: Co-Immunoprecipitation Assay, Western Blot, Expressing, Control, Staining, Confocal Microscopy, Binding Assay, ChIP-qPCR, shRNA, Immunopeptidomics