Journal: Canadian Respiratory Journal

Article Title: Role of Erythromycin-Regulated Histone Deacetylase-2 in Benign Tracheal Stenosis

doi: 10.1155/2020/4213807

Figure Lengend Snippet: The expression of HDAC2 in the different groups determined by immunofluorescence. Image scale bar, 100 µ m. Red fluorescence (Cy3 staining) for the detection of the target protein HDAC2, blue fluorescence (DAPI staining) for the nucleus. Data are presented as the mean ± standard deviation in the corresponding histogram. ∗ P < 0.05 vs. the control group. Control: rabbit tracheal stenosis model without treatment; NS: rabbit tracheal stenosis model treated with penicillin; ERY: rabbit tracheal stenosis model treated with erythromycin; Budesonide: rabbit tracheal stenosis model treated with budesonide; Vorinostat: rabbit tracheal stenosis model treated with vorinostat. HDAC2, histone deacetylase-2; IF, immunofluorescence.

Article Snippet: Erythromycin enteric-coated tablets (H42021990, Yichang Humanwell Pharmaceutical Co., LTD.); Vorinostat Capsules (180509, Beijing Hengrui Kangda Medical Science and Technology Development Co., Ltd.); Budesonide (AstraZeneca 8339000); Rabbit Anti-Collagen III Polyclonal Antibody (bs-10423R, Bioss); Rabbit Anti-Collagen I Polyclonal Antibody (bs-0549R, Bioss); Rabbit Anti-HDAC2 Polyclonal Antibody (bs-1813R, Bioss); Rabbit VEGF ELISA kit (MM-021001); Rabbit TGF- β 1 ELISA kit (MM-3684001); Rabbit Polyclonal Anti-VEGF (bs-1313R, Bioss, 1/500–1/2000); Rabbit Polyclonal Anti-TGF β 1 (bs-0086R, Bioss, 1/500–1/2000); Rabbit monoclonal Anti-IL-8 (ab34100, abcam, 1/1000); Rabbit Polyclonal Anti-HDAC2 (OmnimAbs, OM105905, 1/500–1/2000); fluorescence microscope (CKX53, OLYMPUS); Microplate Reader (RT-6100, Rayto); Protein vertical electrophoresis instrument (DYY-6C, Beijing 61 instrument factory); Ultra High Sensitivity Chemiluminescence Imaging System (Chemi DocTM XRS+, Bio-Rad Shanhhai Laboratories).

Techniques: Expressing, Immunofluorescence, Fluorescence, Staining, Standard Deviation, Histone Deacetylase Assay

Journal: Frontiers in Immunology

Article Title: Redirection of Epithelial Immune Responses by Short-Chain Fatty Acids through Inhibition of Histone Deacetylases

doi: 10.3389/fimmu.2015.00554

Figure Lengend Snippet: Immunomodulatory effects of SCFAs are mimicked by the histone deacetylase inhibitor TSA. (A) HDAC activity in HeLa lysates in the presence of 10 mM Na-butyrate, Na-propionate or 2 μM TSA. (B) Immunoblots of HEK293 and HeLa 57A incubated with 10 mM Na-Bu, 10 mM Na-Pro or 2 μM TSA for 30 min or 5 h analyzed with anti-acetyl-lysine antibody to visualize total histone acetylation and anti-HDAC1, HDAC2, HDAC3, and GAPDH antibodies. (C,D) . Immunomodulatory effects of SCFAs are mimics by histone deacetylase inhibitor TSA. TLR5-NF-κB assays of HEK293 or HeLa 57A expressing TLR5 and an NF-κB-luciferase reporter. Cells were incubated with 10 mM Na-Bu, 10 mM Na-Pro or 2 μM TSA for 30 min followed by stimulation with S. enteriditis flagellin for 5 h. (E) TLR4-NF-κB assays of HEK293 or HeLa 57A expressing TLR4 and an NF-κB-luciferase reporter incubated with SCFA and LPS. Bars depict mean and SEM of three independent experiments. p < 0.05 (*), p < 0.01 (**), p < 0.005 (***).

Article Snippet: Other polyclonal antibodies used were directed against acetyl-H3K9 (Merck, 07-352), acetyl-H4K5 (Merck, 07-327), acetyl-H4K16 (Merck, 07-329), HDAC1 (Abcam, ab109411), HDAC2 (Abcam, ab124974) and HDAC3 (Abcam, ab16047).

Techniques: Histone Deacetylase Assay, Activity Assay, Western Blot, Incubation, Expressing, Luciferase

Journal: Journal of Cellular and Molecular Medicine

Article Title: Histone deacetylase HDAC2 silencing prevents endometriosis by activating the HNF4A/ARID1A axis

doi: 10.1111/jcmm.16835

Figure Lengend Snippet: HDAC2 expression is high in endometriosis. (A) The volcano map of gene expression in GSE37837 . The black dot indicates the genes with no significant difference, the red dot indicates the significantly upregulated gene, and the green dot indicates the significantly downregulated gene. (B) Venn map of 40 intersected genes between the significantly upregulated genes of GSE37837 and the gene retrieved by GeneCards. (C) The importance of genes in endometriosis analysed by phenolyzer. When the length of the column was longer, the importance was higher. (D) HDAC2 expression in GSE37837 . The left blue box showed the expression in normal samples, and the right red box showed the expression in endometriosis samples. The data were expressed as median ± interquartile range. (E) GEPIA of the expression of HDAC2 in UCEC. The left red box was the expression in UCEC, and the right grey box was the expression in the control group. The data were expressed as median ± interquartile range. * p < 0.05. (F) The expression of HDAC2 protein in normal tissues ( n = 30) and endometriosis tissues ( n = 40) detected by immunohistochemistry. The data were expressed as median ± interquartile range. (G) Western blot analysis of the expression of HDAC2 protein in normal tissues ( n = 30) and endometriosis tissues ( n = 40). The data were expressed as median ± interquartile range. * p < 0.05 vs. the control group. The experiment was repeated three times independently

Article Snippet: Primary rabbit anti‐human antibodies to HDAC2 (12922‐3‐AP, 1:200, Proteintech), HNF4A (ab92378, 1:500, Abcam), Ki‐67 (ab16667, 1:200, Abcam), and ARID1A (ab182560, 1:1000, Abcam) were added to the sections for overnight culture at 4°C, and the secondary antibody (ab6785, 1:1000, Abcam) was added.

Techniques: Expressing, Gene Expression, Control, Immunohistochemistry, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Histone deacetylase HDAC2 silencing prevents endometriosis by activating the HNF4A/ARID1A axis

doi: 10.1111/jcmm.16835

Figure Lengend Snippet: Silencing HDAC2 decreases the proliferation and invasion and increases the apoptosis of endometriosis cells. (A) The expression of HDAC2 in hEM15A cells of each group detected by Western blot analysis. (B) The proliferation of hEM15A cells in each group detected by CCK‐8 method. (C) The invasion of hEM15A cells in each group measured by Transwell assay. (D) The apoptosis of hEM15A cells in each group detected by flow cytometry. Data were shown as median ± standard error of mean ( n = 3). * p < 0.05 vs. the sh‐NC group. The experiment was repeated three times independently

Article Snippet: Primary rabbit anti‐human antibodies to HDAC2 (12922‐3‐AP, 1:200, Proteintech), HNF4A (ab92378, 1:500, Abcam), Ki‐67 (ab16667, 1:200, Abcam), and ARID1A (ab182560, 1:1000, Abcam) were added to the sections for overnight culture at 4°C, and the secondary antibody (ab6785, 1:1000, Abcam) was added.

Techniques: Expressing, Western Blot, CCK-8 Assay, Transwell Assay, Flow Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: Histone deacetylase HDAC2 silencing prevents endometriosis by activating the HNF4A/ARID1A axis

doi: 10.1111/jcmm.16835

Figure Lengend Snippet: HDAC2 reduces HNF4A expression through deacetylation. (A) Pearson correlation map of HDAC2 and HNF4A expression in GSE37837 . (B) The expression of HNF4A protein in normal tissues ( n = 30) and endometriosis tissues ( n = 40) determined by immunohistochemistry. The data were expressed as median ± interquartile range. (C) The expression of HNF4A protein in normal tissues ( n = 30) and endometriosis tissues ( n = 40) measured by Western blot analysis. The data were expressed as median ± interquartile range. (D) The enrichment of HDAC2 in the promoter region of HNF4A in normal tissues ( n = 30) and endometriosis tissues ( n = 40) measured by ChIP. (E) The enrichment of HDAC2 in the promoter region of HNF4A of each group detected by ChIP. (F) The expression of HDAC2 and HNF4A in each group assessed by Western blot analysis. (G) HNF4A acetylation level following IP. Data were shown as median ± standard error of mean ( n = 6). * p < 0.05 vs. the sh‐NC group; # p < 0.05 vs. the oe‐NC group. The experiment was repeated three times independently

Article Snippet: Primary rabbit anti‐human antibodies to HDAC2 (12922‐3‐AP, 1:200, Proteintech), HNF4A (ab92378, 1:500, Abcam), Ki‐67 (ab16667, 1:200, Abcam), and ARID1A (ab182560, 1:1000, Abcam) were added to the sections for overnight culture at 4°C, and the secondary antibody (ab6785, 1:1000, Abcam) was added.

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Histone deacetylase HDAC2 silencing prevents endometriosis by activating the HNF4A/ARID1A axis

doi: 10.1111/jcmm.16835

Figure Lengend Snippet: HDAC2 silencing elevates HNF4A expression to slow the proliferation and invasion and facilitate the apoptosis of endometriosis cells. (A) The expression of HDAC2 and HNF4A in hEM15A cells of each group evaluated by Western blot analysis. (B) The viability of hEM15A cells determined by CCK‐8 method. (C) hEM15A cell invasion in each group measured by Transwell assay. (D) Apoptosis of hEM15A cells in each group detected by flow cytometry. Data were shown as median ± standard error of mean ( n = 3). * p < 0.05 vs. the sh‐NC + sh‐NC group; # p < 0.05 vs. the sh‐HDAC2 + sh‐NC group. The experiment was repeated three times independently

Article Snippet: Primary rabbit anti‐human antibodies to HDAC2 (12922‐3‐AP, 1:200, Proteintech), HNF4A (ab92378, 1:500, Abcam), Ki‐67 (ab16667, 1:200, Abcam), and ARID1A (ab182560, 1:1000, Abcam) were added to the sections for overnight culture at 4°C, and the secondary antibody (ab6785, 1:1000, Abcam) was added.

Techniques: Expressing, Western Blot, CCK-8 Assay, Transwell Assay, Flow Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: Histone deacetylase HDAC2 silencing prevents endometriosis by activating the HNF4A/ARID1A axis

doi: 10.1111/jcmm.16835

Figure Lengend Snippet: Endometriosis is repressed in vivo by silencing HDAC2. (A) The area of endometriosis tissues of mice in each group. (B) Endometriosis tissue weight of mice in each group. (C) The expression of HDAC2, HNF4A and ARID1A in endometriosis tissues of each group analysed by immunohistochemistry. (D) Ki‐67 expression measured by immunohistochemistry and apoptosis detected by TUNEL staining in each group. Data were shown as median ± standard error of mean ( n = 8). * p < 0.05 vs. the sh‐NC group. n = 8 mice/group. The experiment was repeated three times independently

Article Snippet: Primary rabbit anti‐human antibodies to HDAC2 (12922‐3‐AP, 1:200, Proteintech), HNF4A (ab92378, 1:500, Abcam), Ki‐67 (ab16667, 1:200, Abcam), and ARID1A (ab182560, 1:1000, Abcam) were added to the sections for overnight culture at 4°C, and the secondary antibody (ab6785, 1:1000, Abcam) was added.

Techniques: In Vivo, Expressing, Immunohistochemistry, TUNEL Assay, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Histone deacetylase HDAC2 silencing prevents endometriosis by activating the HNF4A/ARID1A axis

doi: 10.1111/jcmm.16835

Figure Lengend Snippet: Molecular mechanism of the HDAC2/HNF4A/ARID1A axis involved in endometriosis. HDAC2 inhibited HNF4A through deacetylation, thus diminishing ARID1A expression to promote endometriosis

Article Snippet: Primary rabbit anti‐human antibodies to HDAC2 (12922‐3‐AP, 1:200, Proteintech), HNF4A (ab92378, 1:500, Abcam), Ki‐67 (ab16667, 1:200, Abcam), and ARID1A (ab182560, 1:1000, Abcam) were added to the sections for overnight culture at 4°C, and the secondary antibody (ab6785, 1:1000, Abcam) was added.

Techniques: Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Expression of GR-α and HDAC2 in steroid-Sensitive and steroid-Insensitive interstitial lung disease.

doi: 10.1016/j.biopha.2019.109380

Figure Lengend Snippet: Fig. 3. Expression of HDAC2 in lung tissues in each group. Specificity of polyclonal anti-HDAC2 antibody. Representative expression of (A) HDAC2 protein in lung tissues of controls, (B) COP lung tissues, (C) IPF lung tissues, (D) PA tissues, and (E) pulmonary inflammatory pseudotumor tissues. (F) Mean optical density of HDAC2 in the lungs for each group. Image magnification, ×200 and ×400. Cell positivity is shown by brown staining. *p < 0.05.

Article Snippet: Tissue sections were subsequently incubated overnight at 4 °C with the following primary antibodies: prediluted polyclonal rabbit IgG antibody directed against HDAC2 (1:700, Wuhan Boster Biological Technology, Wuhan, China), monoclonal mouse IgG antibody directed against GR-α (1:700, Cell Signaling Technology, Beverly, MA, USA), monoclonal rabbit IgG antibody directed against NF-κBp65 (1:8000, IPF, idiopathic pulmonary fibrosis; COP, cryptogenic organizing pneumonia; PA, pulmonary abscess; PIP, pulmonary inflammatory pseudotumor; PFT: Pulmonary function test; FEV1: Forced expiratory volume in one second; FVC: forced vital capacity.

Techniques: Expressing, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Expression of GR-α and HDAC2 in steroid-Sensitive and steroid-Insensitive interstitial lung disease.

doi: 10.1016/j.biopha.2019.109380

Figure Lengend Snippet: Fig. 6. Correlation between HDAC2, GR-α, and NF-κBp65 in COP. (A) Correlation between GR-α and NF-κBp65 in COP patients (r = −0.6943, p = 0.038). (B) Correlation between HDAC2 and NF-κBp65 in COP patients (r = −0.8077, p = 0.0085). Correlations between two parameters were determined by Spearman’s rank correlation test.

Article Snippet: Tissue sections were subsequently incubated overnight at 4 °C with the following primary antibodies: prediluted polyclonal rabbit IgG antibody directed against HDAC2 (1:700, Wuhan Boster Biological Technology, Wuhan, China), monoclonal mouse IgG antibody directed against GR-α (1:700, Cell Signaling Technology, Beverly, MA, USA), monoclonal rabbit IgG antibody directed against NF-κBp65 (1:8000, IPF, idiopathic pulmonary fibrosis; COP, cryptogenic organizing pneumonia; PA, pulmonary abscess; PIP, pulmonary inflammatory pseudotumor; PFT: Pulmonary function test; FEV1: Forced expiratory volume in one second; FVC: forced vital capacity.

Techniques:

Journal: Oncotarget

Article Title: The subcellular distribution and function of MTA1 in cancer differentiation

doi:

Figure Lengend Snippet: A, Western blot analysis showing the localization of HDA2 to both the nucleus (N) and cytoplasm (C) of HCT116 cells; B, In vitro immunoprecipitation analysis of the MTA1-HDAC2 interaction in the nuclear and cytoplasmic fractions of HCT116-M1 cells; C, In vitro immunoprecipitation analysis of the MTA1-HDAC2 interaction in the nuclear and cytoplasmic fractions of HCT116-M3 cells; D, In situ PLA analysis. left, the control interaction detected using mouse antibody against MTA1 and rabbit IgG; right, in situ PLA visualization of the MTA1-HDAC2 interaction in both the nucleus and cytoplasm of HCT116 cells. The white arrow shows the positive signal in the cytoplasm.

Article Snippet: The following antibodies were used: mouse monoclonal antibody anti-MTA1 (Abcam); rabbit polyclonal antibody anti-MTA1 (Abcam); mouse monoclonal antibody anti-HDAC2 (Abcam); rabbit polyclonal antibody anti- HDAC2 (Bioworld Technology); rabbit polyclonal antibody anti-GAPDH (Bioworld Technology); rabbit polyclonal antibody anti-histone H3 (Bioworld Technology); and rabbit polyclonal antibody anti-TPR (Santa Cruz).

Techniques: Western Blot, In Vitro, Immunoprecipitation, In Situ