primary antibodies against hdac6  (Proteintech)

Journal: Nutrients

Article Title: An 8-Week Very Low-Calorie Ketogenic Diet (VLCKD) Alters the Landscape of Obese-Derived Small Extracellular Vesicles (sEVs), Redefining Hepatic Cell Phenotypes

doi: 10.3390/nu16234189

Figure Lengend Snippet: Gene expression of PIK3CB, AKT1, HDAC10, HDAC6, HDAC3, MMP19, MMP2, MMP9, ACTA2, and VIM in HEPA–RG cells treated with serum-derived sEVs from obese patients after VLCKD for 8 weeks. Patients with LR and patients with IHR at T0 (before the VLCKD) and after 8 weeks of VLCKD (T1). For the HEPA-RG expression, the fold change values for PIK3CB and AKT1 are reported in ( A ). The fold change values for HDAC 10, HDAC 6, and HDAC 3 are reported in ( B ). The fold change values for MMP19, MMP9, and MMP3 are reported in ( C ). The fold change values for VIM and ACTA2 are reported in ( D ). (* p < 0.05, ** p < 0.001, and *** p < 0.0001 for T1 vs. T0).

Article Snippet: The membranes were incubated with primary antibodies against PIK3CB, AKT, p -AKT, MMP2, MMP9, VIM, HDAC3, and HDAC6 (1:500 dilution, Cell Signaling Technology, Beverly, MA, USA), anti-α-SMA (1:400, Thermo Fisher), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000, Santa Cruz, Santa Cruz, CA, USA).

Techniques: Gene Expression, Derivative Assay, Expressing

Journal: Nutrients

Article Title: An 8-Week Very Low-Calorie Ketogenic Diet (VLCKD) Alters the Landscape of Obese-Derived Small Extracellular Vesicles (sEVs), Redefining Hepatic Cell Phenotypes

doi: 10.3390/nu16234189

Figure Lengend Snippet: Evaluation of proteins regulating hepatocyte degeneration and liver fibrosis was conducted in HEPA-RG cell lines treated with sEVs isolated from patients with LR and patients with IHR, both before (T0) and after (T1) 8 weeks of VLCKD. Representative Western blots of various proteins (PIK3CB, AKT, pAKT, HDAC3, HDAC6, α-SMA, and VIM) and the housekeeping protein GAPDH ( A ). A semi-quantitative evaluation of protein expression levels was performed using video-densitometry analysis of PIK3CB, AKT, and pAKT1 ( B ), HDAC6 and HDAC3 ( C ), MMP2 and MMP9 ( D ), and α-SMA and VIM ( E ) bands on the Western blots. The GAPDH protein band was used to normalize the protein bands for each subject. (* p < 0.05 and ** p < 0.001 for T1 vs. T0).

Article Snippet: The membranes were incubated with primary antibodies against PIK3CB, AKT, p -AKT, MMP2, MMP9, VIM, HDAC3, and HDAC6 (1:500 dilution, Cell Signaling Technology, Beverly, MA, USA), anti-α-SMA (1:400, Thermo Fisher), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000, Santa Cruz, Santa Cruz, CA, USA).

Techniques: Isolation, Western Blot, Expressing

Journal: Cell chemical biology

Article Title: A Cell-based Target Engagement Assay for the Identification of Cereblon E3 Ubiquitin Ligase Ligands and Their Application in HDAC6 Degraders

doi: 10.1016/j.chembiol.2020.04.008

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Primary antibody against HDAC1, HDAC2, HDAC4, HDAC6, IKZF1, IKZF3 and GSPT1 were purchased from Cell Signaling Technology.

Techniques: Recombinant, Software

Journal: Frontiers in Oncology

Article Title: MPT0G413, A Novel HDAC6-Selective Inhibitor, and Bortezomib Synergistically Exert Anti-tumor Activity in Multiple Myeloma Cells

doi: 10.3389/fonc.2019.00249

Figure Lengend Snippet: MPT0G413 potently inhibited histone deacetylase (HDAC6) and inhibited multiple myeloma cell growth and proliferation. (A) Chemical structure of MPT0G413. (B) Human multiple myeloma cell lines (RPMI-8226, NCI-H92; density, 1 × 10 4 ) and human bone marrow stromal cells (HS-5; density, 5 × 10 3 ) were incubated with or without the indicated concentrations of MPT0G413 for 48 h. Cell proliferation was evaluated using a 5-bromo-2′-deoxyuridine (BrdU) proliferation assay. (C) RPMI-8226, NCI-H929, and HS-5 cells were exposed to MPT0G413 at the indicated concentrations for 24, 48, and 72 h. Cell viability was measured using a MTT assay. (D,E) RPMI-8226 and NCI-H929 cells were treated with DMSO or MPT0G413 (0.1, 1, 2.5, 10 μM) and SAHA (2.5 μM) for 24 h. Cells were subsequently harvested, and the lysates were subjected to Western blotting of the indicated proteins. Protein levels in the Western blots were quantified using Image J software. The results are shown as mean ± SEM from three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001, compared with HS-5 cells group (B) and the control group (C–E) .

Article Snippet: We used non-conjugated primary antibodies against HDAC6 (#7612), Caspases-3 (#9661),−8 (#9746), and−9 (#9502), acetyl-histone 3 (#9677), acetyl-histone 4 (#8647), histone 3 (#9715), histone 4 (#2935), acetyl-α-tubulin (#5335), were purchased from Cell Signaling Technology (Danvers, MA, USA). α-tubulin (GTX112141), dynein (GTX80684), ubiquitin (GTX19247), ICAM (GTX100450), LC3B (GTX127375), acetyl-histone 2 (GTX633388) and histone 2 (GTX129418) were purchased from GeneTex (Hsinchu, Taiwan).

Techniques: Histone Deacetylase Assay, Incubation, Proliferation Assay, MTT Assay, Western Blot, Software, Control

Journal: Frontiers in Oncology

Article Title: MPT0G413, A Novel HDAC6-Selective Inhibitor, and Bortezomib Synergistically Exert Anti-tumor Activity in Multiple Myeloma Cells

doi: 10.3389/fonc.2019.00249

Figure Lengend Snippet: MPT0G413 inhibited the binding of histone deacetylase (HDAC6) to dynein and induced the significant accumulation of polyubiquitinated proteins when combined with bortezomib. (A) Hypothetical rationale of proteasome and aggresome pathway inhibition by bortezomib and MPT0G413. (B) RPMI-8226 and NCI-H929 cells were treated with MPT0G413 (1 and 2.5 μM) for 6 h, and total cell lysates were subjected immunoprecipitation with 1 μg of an anti-dynein or anti-ubiquitin antibody, followed by immunoblotting with an anti-HDAC6 antibody. (C) RPMI-8226 and NCI-H929 cells were cultured with MPT0G413 (2.5 μM), bortezomib (2.5 nM), or combined therapy for 12 h. Whole cell lysates were subjected to Western blotting with antibodies specific for ubiquitin and LC3. (D) NCI-H929 cells were treated with MPT0G413 (2.5 μM), bortezomib (2.5 nM), or combination therapy for 6 h and stained by anti-LC3B (green) and anti-ubiquitin (red) antibodies and DAPI (blue) prior to confocal microscopy analysis. Scale bar = 50 μm.

Article Snippet: We used non-conjugated primary antibodies against HDAC6 (#7612), Caspases-3 (#9661),−8 (#9746), and−9 (#9502), acetyl-histone 3 (#9677), acetyl-histone 4 (#8647), histone 3 (#9715), histone 4 (#2935), acetyl-α-tubulin (#5335), were purchased from Cell Signaling Technology (Danvers, MA, USA). α-tubulin (GTX112141), dynein (GTX80684), ubiquitin (GTX19247), ICAM (GTX100450), LC3B (GTX127375), acetyl-histone 2 (GTX633388) and histone 2 (GTX129418) were purchased from GeneTex (Hsinchu, Taiwan).

Techniques: Binding Assay, Histone Deacetylase Assay, Inhibition, Immunoprecipitation, Ubiquitin Proteomics, Western Blot, Cell Culture, Staining, Confocal Microscopy