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Pci 34051

Manufactured by Cayman Chemical
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Sourced in United States, Germany

PCI-34051 is a chemical compound produced by Cayman Chemical. It is commonly used in laboratory settings for research purposes. The core function of PCI-34051 is to serve as a chemical tool for scientific investigations, but no further details on its intended use can be provided in an unbiased and factual manner.

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Lab products found in correlation

Apicidin, by Merck Group (2 mentions) Mocetinostat, by Selleck Chemicals (2 mentions) Mc1568, by Merck Group (2 mentions) Droxinostat, by Selleck Chemicals (2 mentions) Ms 275, by Enzo Life Sciences (2 mentions) Forskolin, by Merck Group (2 mentions) Cobalt chloride, by Merck Group (1 mentions) Mitotracker red cm h2xros, by Merck Group (1 mentions) Tetramethylrhodamine methyl ester perchlorate tmrm, by Merck Group (1 mentions) Mdivi 1, by Merck Group (1 mentions) Calcitriol, by Merck Group (1 mentions) Beta actin ab8227, by Abcam (1 mentions) Protein a agarose beads, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 555 dye, by Thermo Fisher Scientific (1 mentions) Gst column, by GE Healthcare (1 mentions) Poly l lysine (pll), by Merck Group (1 mentions) Streptomycin, by HiMedia (1 mentions) Dulbecco modified eagle medium (dmem), by HiMedia (1 mentions) Fetal bovine serum (fbs), by HiMedia (1 mentions) Propidium iodide, by HiMedia (1 mentions)

11 protocols using pci 34051

1

HDAC8 Inhibition and Temozolomide Treatment in Glioblastoma Cells

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U87, T98G, 293T and HeLa cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin. HeLa cells were transfected using polyethylenimine. To generate stable cell lines, T98G and U87 cells were infected with viruses produced in 293T cells using lentiviral vectors for protein overexpression or knockdown.
U87 and T98G cells were treated with the HDAC8-specific inhibitor PCI34051 (Cayman Chemical Company) according to the indicated concentrations and times. Both U87 and T98G cells were treated with 250 µM temozolomide (TMZ) for 2 days.
Santos-Barriopedro I., Li Y., Bahl S, & Seto E. (2019). HDAC8 affects MGMT levels in glioblastoma cell lines via interaction with the proteasome receptor ADRM1. Genes & Cancer, 10(5-6), 119-133.
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2

Genetic Engineering of HDAC8 Mutants

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The HDAC8 activator TM-2-51 (1-Benzoyl-3-phenyl-2-thiourea), the DRP1 inhibitor Mdivi-1 and cobalt chloride were purchased from Sigma-Aldrich (Oakville, Canada). The HDAC8-specific inhibitor PCI-34051 was purchased from Cayman Chemical (Burlington, Canada). Tetramethylrhodamine methyl ester perchlorate (TMRM) and Mito-Tracker® Red CM-H2X ROS were obtained from Sigma-Aldrich and Invitrogen (Molecular Probes), respectively. Antibodies for mitochondrial voltage-dependent anion channel (VDAC) and p38 mitogen-activated protein kinases (p38) were purchased from Cell Signaling Technologies (Danvers, MA). Antibodies for H3K27ac, pan-histone H3 and HIF-1α were obtained from Active Motif (Carlsbad, CA), Bio Vision (Milpitas, CA) and Novus Biologicals (Littleton, CO), respectively. Antibodies for cytochrome C and DRP1 were obtained from Santa Cruz Biotechnology (Dallas, TX). The HDAC8-EGFP plasmid was prepared as previously described38 (link). The HDAC8-H180R-EGFP plasmid was constructed after cloning the mutant HDAC8 from hHDAC8-6His-pET20b-H180R62 (link), using the PCR primers: 5′-AGATTCTCGAGATGGAAGAACCGGAAGAACC-3′ and 5′-TTCAAGAATTCGAGAACCACGACCTTCGATAACA-3′ and inserting into the pEGFP-N1 vector using the XhoI and EcoR1 restriction enzymes.
Ha S.D., Solomon O., Akbari M., Sener A, & Kim S.O. (2018). Histone deacetylase 8 protects human proximal tubular epithelial cells from hypoxia-mimetic cobalt- and hypoxia/reoxygenation-induced mitochondrial fission and cytotoxicity. Scientific Reports, 8, 11332.
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3

Compound Preparation for Cell Signaling Research

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TSA (Sigma-Aldrich, Schnelldorf, Germany: T1952), MC-1568 (Sigma-Aldrich: M1824), Apicidin (Sigma-Aldrich: A8851), MS-275 (Enzo Life Sciences, Lörrach, Germany: ALX-270-378-M005), PCI-34051 (Cayman Chemical, Ann Arbor, MI, USA: 10444), Mocetinostat (Selleckchem, München, Germany: S1122) and Droxinostat (Selleckchem: S1422) were dissolved in dimethyl sulfoxide (AppliChem, Darmstadt, Germany: A3006). Calcitriol was purchased from Sigma-Aldrich. Human TGFβ1 was purified from platelets according to Werz et al.39 (link)
Ahmad K., Katryniok C., Scholz B., Merkens J., Löscher D., Marschalek R, & Steinhilber D. (2014). Inhibition of class I HDACs abrogates the dominant effect of MLL-AF4 by activation of wild-type MLL. Oncogenesis, 3(11), e127-.
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4

Tubulin Acetylation Regulation by HDAC8

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Forskolin was purchased from Sigma (USA). PCI-34051 and Tubastatin were procured from Cayman chemicals. DMEM, FBS, 1% Penicillin & Streptomycin Antibiotics were obtained from Himedia. Monoclonal antibody of HDAC8 (A-4008) was obtained from Epigenetik, Beta actin ab8227 (Abcam), GAPDH (MA5–15738), Alpha tubulin (B-5-1-2) and anti-acetylated alpha tubulin (Cat no: 322700) were purchased from Thermo scientific Life technologies respectively. Protein A Agarose beads were obtained from Santa Cruz. Poly-L-Lysine (Sigma P8920), 4′, 6-Diamindino-2-phenylindole dichloride DAPI as a nuclear stain (Sigma), Alexa Fluor® 555 Dye (Thermo Fischer Scientific). HDAC8 FLUOR DE LYS fluorometric assay kit (BML-AK518–0001) was purchased from Enzo life sciences. Propidium iodide was purchased from Himedia. GST Column (GE Healthcare 17–5132-01), Custom synthesized peptides of alpha tubulin 33–46 amino acids, Acetylated alpha tubulin peptide, at Lys40 DGQMPSDKTIGGGD and Unacetylated Alpha tubulin peptide DGQMPSDKTIGGGD from SIGMA (USA) (resuspended in MilliQ water).
Vanaja G.R., Ramulu H.G, & Kalle A.M. (2018). Overexpressed HDAC8 in cervical cancer cells shows functional redundancy of tubulin deacetylation with HDAC6. Cell Communication and Signaling : CCS, 16, 20.
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5

Comprehensive Cell Culture Protocol

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RPMI culture medium was purchased from Corning (Corning, NY). Fetal bovine serum (FBS) was purchased from Sigma Chemical Co. (St. Louis, MO). Trypsin, pen/strep antibiotics, and puromycin were purchased from Gibco (Grand Island, NY). Trametinib (MEK inhibitor), Panobinostat (pan-HDAC inhibitor), Pictilisib (PI3K inhibitor), Bosentan Hydrate (EDNRB inhibitor), Verteporfin (YAP inhibitor), Entinostat (HDAC1/2/3 inhibitor), and Tubastatin A (HDAC 6 inhibitor) were purchased from Selleckchem (Houston, TX). PCI-34051 (HDAC8 inhibitor) was purchased from Cayman Chemical (Ann Arbor, MI). Endothelin-3 was purchased from Sigma Chemical Co. (St. Louis, MO). WNT5A was purchased from R&D Systems (Minneapolis, MN, USA). Antibodies for Western Blot and immunochemistry were purchased from Cell Signaling Technology (Danvers, MA), Sigma Chemical Co. (St. Louis, MO), Millipore (Bedford, MA) and Abcam (Cambridge, MA). The phospho-Receptor Tyrosine Kinase and phospho-Kinase array were purchased from R&D Systems (Minneapolis, MN, USA). Opti‐MEM medium, Lipofectamine 2000 and Live/Dead viability stain were purchased from Invitrogen/Life Technologies Corp). siRNA for ROR1/2, IGF-1R and YAP were purchased from Dharmacon RNA Technologies (Lafayette, CO). Nontargeting siRNA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). The Endothelin-3 Assay Kit was purchased from IBL (Takasaki, Japan).
Faião-Flores F., Emmons M.F., Durante M.A., Kinose F., Saha B., Fang B., Koomen J.M., Chellappan S., Maria-Engler S.S., Rix U., Licht J.D., Harbour J.W, & Smalley K.S. (2019). HDAC inhibition enhances the in vivo efficacy of MEK inhibitor therapy in uveal melanoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(18), 5686-5701.
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6

Adult SVZ Neurosphere Proliferation Assay

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Adult SVZ cells were seeded into 24-well culture plates at 1 × 105 cells/mL in proliferation media treated with the selective HDAC8 inhibitor PCI-34051 (Cayman Chemical Company, Ann Arbor, MI, USA) or vehicle control (Dimethyl sulfoxide, DMSO). Neurospheres were allowed to develop for 7 days in an incubator with 5% CO2 in a humidified atmosphere at 37 °C. The average neurosphere diameter was assessed in three 2 mm2 squares in each well.
To assess the proliferation of neurospheres derived from adult SVZs in vitro, neurospheres were plated on chamber slide glasses, incubated for 30 min, and then exposed to EdU (10 μM) for another 30 min at 37 °C, followed by fixation in 4% paraformaldehyde. To detect incorporated EdU (Invitrogen, Waltham, MA, USA, E10187), we used a Click-iT® EdU Imaging Kit (Thermo Fisher Scientific, MA, USA) following the manufacturer’s instructions.
Fukuda M., Fujita Y., Hino Y., Nakao M., Shirahige K, & Yamashita T. (2024). Inhibition of HDAC8 Reduces the Proliferation of Adult Neural Stem Cells in the Subventricular Zone. International Journal of Molecular Sciences, 25(5), 2540.
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7

Histone Deacetylase Inhibitor Protocol

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Calcitriol (17936) and the proteasome inhibitor MG132 (Z-Leu-Leu-Leu-al, C2211) were purchased from Sigma-Aldrich, Schnelldorf, Germany. Human TGFβ1 was purified from platelets according to [41 (link)]. TSA (Sigma-Aldrich: T1952), MC-1568 (Sigma-Aldrich: M1824), Apicidin (Sigma-Aldrich: A8851), MS-275 (Enzo Life Sciences, Lörrach, Germany: ALX-270-378-M005), PCI-34051 (Cayman Chemical, Ann Arbor, MI, USA: 10444), Mocetinostat (Selleckchem, München, Germany: S1122), Droxinostat (Selleckchem: S1422) and Flavopiridol (Sigma-Aldrich: F3055) were all dissolved in DMSO (AppliChem, Darmstadt, Germany: A3006). AFF1 ELISA Kit (E98872Hu) was obtained from USCN Life Science, Wuhan, Hubei, PRC.
Ahmad K., Scholz B., Capelo R., Schweighöfer I., Kahnt A.S., Marschalek R, & Steinhilber D. (2015). AF4 and AF4-MLL mediate transcriptional elongation of 5-lipoxygenase mRNA by 1, 25-dihydroxyvitamin D3. Oncotarget, 6(28), 25784-25800.
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8

Inhibition of Epigenetic Regulators

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PA and LF were purchased from the List Biological Laboratories (Campbell, CA, USA). Chemical inhibitors used in this study are the following: PCI-34051(Cayman chemical, Ann Arbor, MI, USA), TM-2-51(1-Benzoyl-3-phenyl-2-thiourea, Sigma-Aldrich, St.Louis, MO, USA), JMJD3 inhibitor GSK-J4 (Sigma-Aldrich), EZH2 inhibitor EPZ-6438 (MedChemexpress CO., Ltd; Princeton, NJ, USA, through CEDARLANE), PI3K inhibitors LY294002 (ApexBio Technology, Houston, TX, USA) and wortmannin (Calbiochem, La Jolla, CA, USA), PTEN inhibitor VO-OHpic trihydrate (BioVision, Milpitas, CA, USA). Propidium iodide and RNase A were obtained from Calbiochem and Sigma-Aldrich.
Antibodies raised against the NH2-terminus of MEK1 (MEK1-NT) and MEK3 were obtained from Stressgen Bioreagents (Ann Arbor, MI, USA) and Santa Cruz Biotechnology (Dallas, TX, USA), respectively. The phospho-AKT (Ser-473) and PTEN antibodies were purchased from Cell Signaling and Cedarlane (Danvers, MA, USA and Burlington, NC, USA). Anti- H3K27Ac and anti-H3K27me3 antibodies were from Active Motif (Carlsbad, CA, USA); pan-histone H3 antibodies from Bio Vision; β-actin antibodies from Rockland Inc (Gilbertsville, PA, USA).
Ha S.D., Cho W, & Kim S.O. (2017). HDAC8 Prevents Anthrax Lethal Toxin-induced Cell Cycle Arrest through Silencing PTEN in Human Monocytic THP-1 Cells. Toxins, 9(5), 162.
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9

Chemical Reagents and Suppliers for Research

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All chemicals were obtained from Nacalai Tesque (Nacalai Tesque, Kyoto, Japan), Wako (Wako Pure Chemicals, Osaka, Japan), Corning (Corning, Corning, NY, USA), Qiagen (Qiagen, Hilden, Germany), Invitrogen (Invitrogen, Carlsbad, CA, USA), and Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA). TM251 was purchased from Active Motif (Active Motif, Tokyo, Japan). PCI34051, and RGFP966 were acquired from Cayman (Cayman Chemical, Ann Arbor, MI, USA).
Yuliana A., Jheng H.F., Kawarasaki S., Nomura W., Takahashi H., Ara T., Kawada T, & Goto T. (2018). β-adrenergic Receptor Stimulation Revealed a Novel Regulatory Pathway via Suppressing Histone Deacetylase 3 to Induce Uncoupling Protein 1 Expression in Mice Beige Adipocyte. International Journal of Molecular Sciences, 19(8), 2436.
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10

Histone Deacetylase Inhibition in Cell Treatment

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Three cycles of 48-h treatment with PCI34051 (HDAC8 inhibitor, Cayman chemical, cat. no 10444) or TMP 195 (HDAC4 and HDAC7 inhibitor, Cayman chemical, cat. no 23242) were performed. 24 h after seeding, the cells were treated with one first cycle of inhibition. Chemotherapy treatment was performed together with the second cycle of inhibition.
Pantelaiou-Prokaki G., Mieczkowska I., Schmidt G.E., Fritzsche S., Prokakis E., Gallwas J, & Wegwitz F. (2022). HDAC8 suppresses the epithelial phenotype and promotes EMT in chemotherapy-treated basal-like breast cancer. Clinical Epigenetics, 14, 7.
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