hdac5  (Santa Cruz Biotechnology)

Journal: Biomaterials Research

Article Title: HDAC5-mediated exosomal Maspin and miR-151a-3p as biomarkers for enhancing radiation treatment sensitivity in hepatocellular carcinoma

doi: 10.1186/s40824-023-00467-7

Figure Lengend Snippet: Modulation of protein p53 and HDAC5 expression in HCC after RT. Following radiation exposure at 2 Gy and 4 Gy, protein expression of p53, P21, and HDAC5 were confirmed by western blot in a time-dependent manner up to 24 h per 6 h ( A-F ). The expression of each protein was quantified based on b-actin levels. Data are shown as mean ± SEM of three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. untreated by Student’s t-test

Article Snippet: Pre-designed HDAC5 siRNA and Scrambled siRNA / miRNA controls were purchased from BIONEER (Seoul).

Techniques: Expressing, Western Blot

Journal: Biomaterials Research

Article Title: HDAC5-mediated exosomal Maspin and miR-151a-3p as biomarkers for enhancing radiation treatment sensitivity in hepatocellular carcinoma

doi: 10.1186/s40824-023-00467-7

Figure Lengend Snippet: Interaction between HDAC5 and p53 induced by RT and activation of HDAC5 mediated p53-dependent exosome secretion pathway in HCC. RT-induced expression of HDAC5, p53, P21, and PUMA in p53 −/− and p53 +/+ HepG2 was confirmed by western blot and quantified using b-actin as an internal control ( A ). Data are shown as mean ± SEM of three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. untreated by Student’s t-test. The interaction between HDAC5 and p53 after RT was confirmed using Duolink proximity ligation assay in HepG2 ( B ). p53 +/+ HepG2 cells were treated with 4 Gy radiation and incubated for 12 h, and were then immunoprecipitated with anti-p53 antibodies. Precipitated proteins were subjected to co-immunoprecipitation (CO-IP) using an anti-HDAC5 antibody and analyzed by immunoblotting ( C ). The expression of TSAP6 induced by RT in p53 −/− and p53 +/+ HepG2 was verified by western blot. TSAP6 expression was quantified using b-actin as an internal control ( D ). Data are shown as mean ± SEM of three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. untreated by Student’s t-test. Twenty-four hours after siHDAC5 was transfected into HepG2, exposed to 4 Gy radiation. After 12 h incubation, HDAC5, p53, and TSAP6 expressions were confirmed by western blot and quantified using b-actin as an internal control ( E ). Data are shown as mean ± SEM of three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. untreated by Student’s t-test; # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. 4 Gy treatment by Student’s t-test

Article Snippet: Pre-designed HDAC5 siRNA and Scrambled siRNA / miRNA controls were purchased from BIONEER (Seoul).

Techniques: Activation Assay, Expressing, Western Blot, Proximity Ligation Assay, Incubation, Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection

Journal: Biomaterials Research

Article Title: HDAC5-mediated exosomal Maspin and miR-151a-3p as biomarkers for enhancing radiation treatment sensitivity in hepatocellular carcinoma

doi: 10.1186/s40824-023-00467-7

Figure Lengend Snippet: Expression of upstream regulator HDAC5-dependently regulated cellular and exosomal Maspin and anti-angiogenic function caused by human recombinant Maspin. HDAC5 siRNA was transfected into HepG2 in a dose-dependent manner, and after 24 h, HDAC5, p53, Maspin, and TSAP6 expressions were analyzed by western blot. The expression of each protein was quantified using b-actin as an internal control ( A ). After HDAC5 siRNA was transfected into HUVECs, Maspin mRNA expression was investigated. It was confirmed using the GSE15499 GEO data set ( B ). Data are shown as mean ± SD of three independent values. * P < 0.05 vs. con siRNA by Student’s t-test. HepG2 was exposed to 4 Gy RT 24 h after HDAC5 siRNA was transfected. After overnight, the expression of HDAC5, Maspin, HSP90, and Calnexin in cell lysates and exosomes of each group was examined by western blot ( C , D ). HUVECs were seeded on growth factor-reduced matrigel-coated plates and treated with VEGF (50 ng/ml), human recombinant Maspin (1-10ug/ml). After 16 h, calcein am dye (0.5ug/ml) was treated and fluorescence images were photographed using EVO M5000 Imaging System (Invitrogen) ( E ). Five locations of each well were randomly photographed, and the branching and tube organization were measured and compared with that of the control condition ( F , G ). Cell culture dishes were performed in at least three separate experiments. Scale bar = 200 μm. Data are shown as mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control by Student’s t-test

Article Snippet: Pre-designed HDAC5 siRNA and Scrambled siRNA / miRNA controls were purchased from BIONEER (Seoul).

Techniques: Expressing, Recombinant, Transfection, Western Blot, Fluorescence, Imaging, Cell Culture

Journal: Biomaterials Research

Article Title: HDAC5-mediated exosomal Maspin and miR-151a-3p as biomarkers for enhancing radiation treatment sensitivity in hepatocellular carcinoma

doi: 10.1186/s40824-023-00467-7

Figure Lengend Snippet: Exosomal Maspin and miR-151a-3p: Biomarkers Enhancing Radiation Treatment Sensitivity in HCC. A schematic image; In HCC, RT can alter exosomal components by mediating HDAC5 and p53, and exosomal Maspin and miR-151a-3p have the potential as biomarkers that can modulate radiosensitivity

Article Snippet: Pre-designed HDAC5 siRNA and Scrambled siRNA / miRNA controls were purchased from BIONEER (Seoul).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Curcuphenol possesses an unusual histone deacetylase enhancing activity that counters immune escape in metastatic tumours

doi: 10.3389/fphar.2023.1119620

Figure Lengend Snippet: Primer sequences used for RT-qPCR.

Article Snippet: TC1 cells were transfected with 7 μL of siRNAs HDAC5, HDAC8, HDAC10 or Control siRNA-A (Santa Cruz).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Stenotrophomonas maltophilia promotes lung adenocarcinoma progression by upregulating histone deacetylase 5

doi: 10.3389/fmicb.2023.1121863

Figure Lengend Snippet: Stenotrophomonas maltophilia promoted cell and migration in lung epithelial cells. (A) Schematic experimental design for two in vitro experiments. For all conditions, A549 cells were exposed for 4 h and then harvested for RNA isolation. Experiment 1: Cells were infected with S . maltophilia (ATCC #13637) at three different multiplicity of infection (MOI) of 1, 10, and 100 for 4 h. Experiment 2: exposure to media alone, S . maltophilia (heat-killed), and supernatant. (B) Effect of S . maltophilia on A549 cell viability. (C) Effect of heat-killed S . maltophilia and supernatant on A549 cell viability, ns: no significance. (D) Effects of S . maltophilia on A549 cell line migration. (E) Effects of S . maltophilia on HDAC5 gene expression in A549 cells. ** p < 0.01 compared with CTL, *** p < 0.001 compared with CTL. (F) Western blot and statistical analysis of HDAC5 expression in A549 cells. * p < 0.05. ns, no significance.

Article Snippet: When cells reached 60–80% confluence, the cells were transfected with HDAC5 siRNA, and control vector (Guangzhou RiboBio Co., Ltd., Guangzhou, China) using LipoRNAiTM Transfection Reagent (Beyotime, #C0535, Shanghai, China) according to the manufacturer’s specification.

Techniques: Migration, In Vitro, Isolation, Infection, Gene Expression, Western Blot, Expressing

Journal: Frontiers in Microbiology

Article Title: Stenotrophomonas maltophilia promotes lung adenocarcinoma progression by upregulating histone deacetylase 5

doi: 10.3389/fmicb.2023.1121863

Figure Lengend Snippet: Inhibition of HDAC5 suppressed lung cancer cellular functions. Lung cancer cells A549 were transfected with si-HDAC5 or negative control (si-NC). (A) The HDAC5 expression levels were detected by qRT-PCR. (B) CCK-8 assay was used to test the cell proliferation. (C) Transwell migration assay was used to test the cell migration ability. * p < 0.05, *** p < 0.001 by Student’s t-test.

Article Snippet: When cells reached 60–80% confluence, the cells were transfected with HDAC5 siRNA, and control vector (Guangzhou RiboBio Co., Ltd., Guangzhou, China) using LipoRNAiTM Transfection Reagent (Beyotime, #C0535, Shanghai, China) according to the manufacturer’s specification.

Techniques: Inhibition, Transfection, Negative Control, Expressing, Quantitative RT-PCR, CCK-8 Assay, Transwell Migration Assay, Migration