Journal: Genes & Development

Article Title: Epe1 recruits BET family bromodomain protein Bdf2 to establish heterochromatin boundaries

doi: 10.1101/gad.221010.113

Figure Lengend Snippet: Bdf2 counteracts Sir2-mediated histone deacetylation. ( A , C ) Serial dilution analysis to measure heterochromatin spreading outside of IRC1R ∷ ura4 + . ( B , D , E ) ChIP analysis of H3K9me2 and Swi6 levels at IRC1R∷ura4 + and H4K16ac levels at IRC1 , normalized to act1 . The data are averages of three experiments, and error bars represent standard deviation. ( F , left ) Coomassie-stained gel of recombinant Sir2. ( Right ) HDAC assays were performed with recombinant Sir2, a tetra-acetylated histone H4 peptide, and recombinant Bdf2. The production of nicotinamide was measured via the generation of nicotinic acid and free ammonia by nicotinamidase.

Article Snippet: Sir2 HDAC assay was performed using the SIRTainty Class III HDAC assay kit (Millipore) according to the manufacturer's protocol.

Techniques: Serial Dilution, Standard Deviation, Staining, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: Histone Deacetylase Inhibitors Downregulate Calcium Pyrophosphate Crystal Formation in Human Articular Chondrocytes

doi: 10.3390/ijms23052604

Figure Lengend Snippet: Histone deacetylase inhibitors (HDACis) downregulated class II HDAC member expressions and upregulated the acetylation of histones 3 and 4 in transforming growth factor (TGF)-β1-treated chondrocytes. ( A , B ) Human primary chondrocytes (HC-a cells) were pretreated with different concentrations of trichostatin A (TSA) (100~800 nM) and vorinostat (SAHA) (100–1000 nM) for 30 min and then treated with 5 ng/mL TGF-β1 for 24 h. Total cellular proteins were collected, and protein expressions were detected by Western blotting. Ac, acetylated.

Article Snippet: Two kinds of class I and II HDACis, TSA and SAHA, were obtained from Union Biomed (Taipei, Taiwan); recombinant human TGF-β1 was obtained from R&D Systems (Minneapolis, MN, USA); the anti-ANKH and anti-α-tubulin antibodies were obtained from Novus Biologicals (Littleton, CO, USA); the class I HDAC antibody sampler kit, class II HDAC antibody sampler kit, anti-Smad2, anti-phospho-Smad2 (Ser465/467), anti-histone H3, anti-acetyl-histone H3-Lys9, and anti-acetylated-lysine (Ac-K-103) antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA); the anti-ENPP1, anti-TNAP, anti-acetyl-histone H4, anti-TGF-β receptor I (TGFBR1), and anti-TGF-β receptor II (TGFBR2) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); and the anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH), anti-mouse immunoglobulin G (IgG) horseradish peroxidase (HRP), and anti-rabbit IgG HRP antibodies were obtained from GeneTex International (Hsinchu City, Taiwan).

Techniques: Histone Deacetylase Assay, Western Blot