Journal: International Journal of Molecular Sciences

Article Title: Class I-Histone Deacetylase (HDAC) Inhibition is Superior to pan-HDAC Inhibition in Modulating Cisplatin Potency in High Grade Serous Ovarian Cancer Cell Lines

doi: 10.3390/ijms20123052

Figure Lengend Snippet: IC 50 and inhibitory constants (K i ) of nexturastat A on HDAC2, 4, 6, and 8.

Article Snippet: Blots were incubated with primary antibodies against acetylated α-tubulin (Cat. No. sc-23950), α-tubulin (Cat. No. sc-8035) (Santa Cruz Biotechnology, Heidelberg, Germany), histone H3 (Cat. No. MAB9448), acetyl histone H3 (Lys24, Cat. No. NBP2-54615), HDAC1 (Cat. No. NB100-56340), HDAC2 (Cat. No. MAB7679), HDAC3 (Cat. No. NB100-1669), HDAC4 (Cat. No. AF6205), HDAC5 (Cat. No. NBP2-22152), HDAC6 (Cat. No. NB100-56343), HDAC10 (Cat. No. NB100-91801), and HDAC11 (Cat. No. NBP2-16789) (Biotechne, Wiesbaden, Germany).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Class I-Histone Deacetylase (HDAC) Inhibition is Superior to pan-HDAC Inhibition in Modulating Cisplatin Potency in High Grade Serous Ovarian Cancer Cell Lines

doi: 10.3390/ijms20123052

Figure Lengend Snippet: Primer sequences for RT-PCR.

Article Snippet: Blots were incubated with primary antibodies against acetylated α-tubulin (Cat. No. sc-23950), α-tubulin (Cat. No. sc-8035) (Santa Cruz Biotechnology, Heidelberg, Germany), histone H3 (Cat. No. MAB9448), acetyl histone H3 (Lys24, Cat. No. NBP2-54615), HDAC1 (Cat. No. NB100-56340), HDAC2 (Cat. No. MAB7679), HDAC3 (Cat. No. NB100-1669), HDAC4 (Cat. No. AF6205), HDAC5 (Cat. No. NBP2-22152), HDAC6 (Cat. No. NB100-56343), HDAC10 (Cat. No. NB100-91801), and HDAC11 (Cat. No. NBP2-16789) (Biotechne, Wiesbaden, Germany).

Techniques:

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: HDAC4 is responsible for deacetylation of GAC at K311. A, B Indicated plasmids were transfected into H1299 cells. Interaction between HDAC4 and GAC was detected by immunoprecipitation and western blot. WCL: whole cell lysate. C Indicated plasmids were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. D Indicated siRNAs were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. E Indicated plasmids and siRNAs were transfected into H1299 cells and glutaminase activity assay was performed. F The mitochondrial and cytosolic proteins in H1299 cells were separated and the location of HDAC4 and GAC was determined by western blot. VDAC was used as a marker of mitochondrial proteins and GAPDH was used as a marker of cytosolic proteins. Data are showed as mean ± SD, n=3. ** P < 0.01, ns P >0.05.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Transfection, Immunoprecipitation, Western Blot, Expressing, Activity Assay, Marker

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: HDAC4 promoted cell proliferation and migration in NSCLC cells. A, B Indicated plasmids were transfected into H1299 (A) and A549 (B) cells and cell growth assay was performed. C, D Indicated siRNAs were transfected into H1299 (C) and A549 (D) cells and cell growth assay was performed. E, F Indicated siRNAs and plasmids were transfected into H1299 (E) and A549 (F) cells and cell growth assay was performed. Western blot assay was performed to confirm the transfection efficiency. G, H Indicated siRNAs were transfected into H1299 cells (G) and H292 cells (H) and cell wound healing assay was performed (scale bar: 500 µm, magnification: 100×). I F-actin staining assay. Indicated siRNAs were transfected into H1299 cells. 48 h later, cells were stained with phalloidin and DAPI. Scale bar=20 µm. Data are showed as mean ± SD, n=3. **P<0.01, ***P<0.001.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Migration, Transfection, Growth Assay, Western Blot, Wound Healing Assay, Staining

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: A working model of GAC acetylation in NSCLC. A working model depicting the molecular mechanism of HDAC4 mediated GAC deacetylation and TRIM21 mediated GAC ubiquitination to regulation tumorigenesis in NSCLC.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques:

Journal: BMC Cancer

Article Title: Functional role of miR-10b in tamoxifen resistance of ER-positive breast cancer cells through down-regulation of HDAC4

doi: 10.1186/s12885-015-1561-x

Figure Lengend Snippet: HDAC4 is a target of miR-10b. a Ingenuity Pathway Analysis for targets of miR-10b. HDAC4 is shown with an arrow. b Sequence complementarities of miR-10b and its target HDAC4. c Luciferase assay was conducted to confirm that HDAC4 is a direct target of miR-10b. MCF-7 cells were co-transfected with dual luciferase plasmid pEZX-MT05-HDAC4-3′UTR along with a control pre-miR or pre-miR-10b, and assayed for luciferase activity 48 h after transfection. d Levels of HDAC4 in parental (MCF-7) and tamoxifen-resistance MCF-7 (MCF7TR) cells. e Effect of altered miR-10b levels on HDAC4 levels. β-actin protein was used as protein loading control for Western blots and RNU48 was used as internal control for the real-time RT-PCR miRNA analysis. C , control; PM , pre-miR-10b; AM , anti-miR-10b

Article Snippet: For luciferase reporter assays, MCF-7 cells were co-transfected with HDAC4 3′UTR luciferase vector (GeneCopoeia, Catalog # HmiT023167-MT05) and pre-miR-10b or miRNA negative control, using DharmaFECT Duo Transfection Reagent (Dharmacon).

Techniques: Sequencing, Luciferase, Transfection, Plasmid Preparation, Control, Activity Assay, Western Blot, Quantitative RT-PCR

Journal: BMC Cancer

Article Title: Functional role of miR-10b in tamoxifen resistance of ER-positive breast cancer cells through down-regulation of HDAC4

doi: 10.1186/s12885-015-1561-x

Figure Lengend Snippet: Effect of HDAC4 levels on tamoxifen-sensitivity. a siRNA against HDAC4 reduced its expression in MCF-7 cells. Functional role of HDAC4 and miR-10b on tamoxifen sensitivity in b MCF-7 and c MCF7TR cells. β-actin protein was used as protein loading control for Western blots. Tamoxifen treatment was done for 48 h at indicated doses. PM , pre-miR-10b; AM , anti-miR-10b; HDAC4 , HDAC cDNA; siHDAC4 , siRNA against HDAC4

Article Snippet: For luciferase reporter assays, MCF-7 cells were co-transfected with HDAC4 3′UTR luciferase vector (GeneCopoeia, Catalog # HmiT023167-MT05) and pre-miR-10b or miRNA negative control, using DharmaFECT Duo Transfection Reagent (Dharmacon).

Techniques: Expressing, Functional Assay, Control, Western Blot

Journal: BMC Cancer

Article Title: Functional role of miR-10b in tamoxifen resistance of ER-positive breast cancer cells through down-regulation of HDAC4

doi: 10.1186/s12885-015-1561-x

Figure Lengend Snippet: miR-10b and its target HDAC4 influence tamoxifen-induced apoptosis and cell viability. Effect of ectopic expression of miR-10b and HDAC4 on ( a ) apoptosis-induction and ( b ) viability of MCF-7 cells, and the effect of silencing of miR-10b and HDAC4 on ( c ) apoptosis-induction and ( d ) viability of MCF7TR cells. Tamoxifen treatment was for 48 h. PM , pre-miR-10b; AM , anti-miR-10b; HDAC4 , HDAC cDNA; siHDAC4 , siRNA against HDAC4

Article Snippet: For luciferase reporter assays, MCF-7 cells were co-transfected with HDAC4 3′UTR luciferase vector (GeneCopoeia, Catalog # HmiT023167-MT05) and pre-miR-10b or miRNA negative control, using DharmaFECT Duo Transfection Reagent (Dharmacon).

Techniques: Expressing

Journal: Biomolecules

Article Title: Participation of the miR-22-HDAC4-DLCO Axis in Patients with COPD by Tobacco and Biomass

doi: 10.3390/biom9120837

Figure Lengend Snippet: Serum HDAC4 protein is higher in COPD by biomass than COPD by smoking. The protein HDAC4 level was quantified in serum by ELISA and was expressed in pg /mL ( n = 25). * p < 0.01. Abbreviations: COPD-BS, COPD biomass smoke exposure; COPD-TS, COPD by tobacco smoke exposure, HDAC4; Histone Deacetylase 4.

Article Snippet: The serum concentration of the human protein histone deacetylase 4 (HDAC4), was assessed using the ELISA test kit (Human Histone Deacetylase 4/HDAC4; EKU04816; Biomatik, Wilmington, DE, USA), following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Histone Deacetylase Assay