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Suberoylanilide hydroxamic acid saha

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Suberoylanilide hydroxamic acid (SAHA) is a synthetic compound used in laboratory research. It functions as a histone deacetylase (HDAC) inhibitor, which can potentially influence gene expression and cellular processes.

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Lab products found in correlation

Vorinostat, by Selleck Chemicals (3 mentions) Furosemide, by Pfizer (2 mentions) Kanamycin, by Merck Group (2 mentions) Tamoxifen, by Merck Group (2 mentions) Bortezomib, by Selleck Chemicals (2 mentions) Tnf α, by Merck Group (1 mentions) Hif 1α shrna lentiviral particles sc 35561 v, by Santa Cruz Biotechnology (1 mentions) Mouse anti human vdr antibody, by Santa Cruz Biotechnology (1 mentions) Etoposide, by MedChemExpress (1 mentions) Doxorubicin dox, by MedChemExpress (1 mentions) Mg132, by Beyotime (1 mentions) Deferoxamine mesylate dfo, by Merck Group (1 mentions) 5 azacytidine aza, by Selleck Chemicals (1 mentions) D erythros phingosine 1 phosphate s1p, by Avanti Polar Lipids (1 mentions) Epidermal growth factor (egf), by R&D Systems (1 mentions) Fulvestrant, by Selleck Chemicals (1 mentions) Tamoxifen, by Selleck Chemicals (1 mentions) Trichostatin a, by Merck Group (1 mentions) Ms 275, by Selleck Chemicals (1 mentions) Lbh 589 panobinostat, by Selleck Chemicals (1 mentions)

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Suberoylanilide hydroxamic acid (4 citations)

11 protocols using suberoylanilide hydroxamic acid saha

1

Generation of Latent HIV-1 Infected Haploid Cell Lines

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We used a strategy described previously (56 (link), 57 (link)) to generate latent HIV-1 infected KBM7 haploid cell lines. Minimal HIV-1 (LTR-GFP, HIV-1-658) (65 (link)) virus was produced by transfection of 293T cells in 15-cm culture dishes using polyethylenimine (PEI) (Sigma-Aldrich) with a mixture of 6.8 μg p658, 2 μg vesicular stomatitis virus G (VSVG), and 4.5 μg Gag-Pol plasmids. Haploid KBM7 cells were infected with HIV-1-derived virus at low MOI such that approximately 5 to 10% of cells became productively infected as determined by GFP expression. Five days after infection, the GFP-negative cell population harboring uninfected and potentially latently infected cells was sorted by flow cytometry-activated cell sorting (FACS) and stimulated with 350 nM suberoylanilide hydroxamic acid (SAHA) (Selleck Chem), and 5 ng/μl TNF-α (Sigma-Aldrich). Twenty-four hours poststimulation, GFP-positive cells were single cell sorted by FACS into 96-well plates. Clones were expanded and characterized for their basal GFP expression and their potential for reactivation. From the clonal cell lines generated, Hap-Lat was selected for low background and relative high reactivation upon stimulation (GFP negative under basal conditions but able to be induced to express GFP upon activation).
Röling M., Mollapour Sisakht M., Ne E., Moulos P., Crespo R., Stoszko M., De Crignis E., Bodmer H., Kan T.W., Akbarzadeh M., Harokopos V., Hatzis P., Palstra R.J, & Mahmoudi T. (2021). A Two-Color Haploid Genetic Screen Identifies Novel Host Factors Involved in HIV-1 Latency. mBio, 12(6), e02980-21.
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2

Characterization of Stem Cell Lineages

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Most immunofluorescence antibodies were purchased from Santa Cruz (Dallas, TX), including rabbit anti-HIF1 and anti-OCT4 antibodies OCT4 monoclonal mouse anti-human antibody, mouse monoclonal anti-human SOX2 antibody, mouse anti-human FOXJ1 monoclonal antibody, mouse anti-human P63 monoclonal antibody, goat anti-human Nanog polyclonal antibody, goat anti-human CCSP polyclonal antibody, rabbit polyclonal anti-human SP-C antibody, mouse anti-human VDR antibody, and secondary rhodamine-labeled and fluorescein-labeled donkey anti-mouse, anti-goat, and anti-rabbit IgG antibodies. TRITC-labeled donkey anti-rabbit IgG antibody was purchased from Thermo Fisher Scientific (Waltham, MA). Several chemicals were used to induce stem cell differentiation, including 1,25(OH)2D3 (VD3), purchased from Sigma and Suberoylanilide hydroxamic acid (SAHA), purchased from Selleck (Houston, TX). HIF-1α shRNA lentiviral particles sc-35561-v was bought from Santa Cruz Biotechnology (Dallas, TX).
Li R., Huang J., Ma M., Lou Y., Zhang Y., Wu L., Chang D.W., Zhao P., Dong Q., Wu X, & Han B. (2016). Two-stage induced differentiation of OCT4+/Nanog+ stem-like cells in lung adenocarcinoma. Oncotarget, 7(42), 68360-68370.
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3

Apoptosis Induction in Cancer Cells

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Doxorubicin (Dox, HY‐15142; MedChem Express, Monmouth Junction, NJ, USA), etoposide (HY‐13629; MedChem Express), nutlin‐3a (SC4368; Beyotime, Shanghai, China), MG132 (S1748; Beyotime), cycloheximide (CHX, 2112S; CST, Beverly, MA, USA), suberoylanilide hydroxamic acid (SAHA, S1047; Selleck Chemicals, Houston, TX, USA) were used. PBS was used as vehicle control for Dox, and DMSO as control for etoposide, nutlin‐3a, SAHA, CHX, and MG132.
Zhang L., Yang J., Luo Y., Liu F., Yuan Y, & Zhuang S. (2020). A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance. Advanced Science, 7(21), 2001364.
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4

Exploring Epigenetic and Signaling Modulators

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Using dimethylsulfoxide (DMSO) treated cells as controls, cells were treated with Temsirolimus (TEMS, #50-811-7, Fisher Scientific, Pittsburgh, PA) at a final concentration of 2μM, as described in our prior work (17 (link)). Likewise, as earlier described (24 (link)), 5-Azacytidine (AZA, #S1782, Selleck Chemicals, Houston, TX) was utilized at a final concentration of 1μM while Suberoylanilide hydroxamic acid (SAHA, #S1047, Selleck Chemicals, Houston, TX) was utilized at a final concentration of 50μM. Lysophosphatidic acid (LPA, as a sodium salt in chloroform, #857130C, Avanti Polar Lipids, Alabaster, AL) was utilized at a final dose of 10μM, as previously described (17 (link)). D-erythro-Sphingosine-1-Phosphate (S1P, #860492, Avanti Polar Lipids, Alabaster, AL) was utilized at a final concentration of 250nM. Deferoxamine mesylate (DFO, #D9533, Sigma-Aldrich, St. Louis, MO) was utilized at a final concentration of 10μM, as described in our earlier work (25 (link)).
Chhabra R., Guergues J., Wohlfahrt J., Rockfield S., Espinoza Gonzalez P., Rego S., Park M.A., Berglund A.E., Stevens SM J.r, & Nanjundan M. (2023). Deregulated expression of the 14q32 miRNA cluster in clear cell renal cancer cells. Frontiers in Oncology, 13, 1048419.
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5

SAHA Drug Delivery Protocol

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Suberoylanilide hydroxamic acid (SAHA) was obtained from Selleck Chemical (Houston, TX, USA). SAHA was dissolved in DMSO at 50 mg/ml to be delivered in a volume that would bypass the potential detrimental effects of excessive DMSO, as described previously.40 (link) Kanamycin (Sigma, St. Louis, MO, USA) was dissolved in 0.9% saline (Hospira, Lake Forest, IL, USA) and delivered by subcutaneous injection. Furosemide (Hospira) was administered by intraperitoneal injection. Tamoxifen (Sigma) was dissolved in corn oil at 25 mg/ml and delivered by intraperitoneal injection at 0.25 mg/g.
Layman W.S., Williams D.M., Dearman J.A., Sauceda M.A, & Zuo J. (2015). Histone deacetylase inhibition protects hearing against acute ototoxicity by activating the Nf-κB pathway. Cell Death Discovery, 1, 15012-.
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6

Synthesis and Characterization of Chemical Compounds

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WT161 (C27H30N4O3, MW = 458.55, Supplementary Figure 1) was synthesized by Dr. Bradner's laboratory. MAZ1793 (Figure 7) was generated by Dr. Mazitschek's laboratory. MS275 (entinostat), suberoylanilide hydroxamic acid (SAHA, vorinostat), LBH589 (panobinostat), CUDC-101, tamoxifen, fulvestrant and bortezomib were purchased from Selleck Chemicals (Houston, TX). Erlotinib was purified from discarded patient tablets. MG132 and trichostatin A (TSA) were obtained from Sigma (St. Louis, MO). Z-VAD-FMK and 17-demethoxygeldanamycin (17-AAG) were purchased from EMD Chemicals (San Diego, CA). Epidermal growth factor was obtained from R&D Systems (Minneapolis, MN).
Hideshima T., Mazitschek R., Qi J., Mimura N., Tseng J.C., Kung A.L., Bradner J.E, & Anderson K.C. (2017). HDAC6 inhibitor WT161 downregulates growth factor receptors in breast cancer. Oncotarget, 8(46), 80109-80123.
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7

SAHA Otoprotection Against Noise Exposure

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Mice were injected intraperitoneally with suberoylanilide hydroxamic acid (SAHA) (Selleckchem, TX), (100 mg/kg body weight) dissolved in DMSO (MilliporeSigma, MA), or with DMSO alone (vehicle) 3 days before exposure to noise and 2 h after the end of the noise exposure. The SAHA dosing amount was based on a previous publication using SAHA as an otoprotective agent, where the authors tried different dose concentrations of SAHA and reported a 100 mg/kg dose to be most efficacious without cytotoxicity [12 (link)]. Because studies vary in their dosing regimen for SAHA, the frequency of the dosing was based on the published literature with minor modifications [14 (link), 31 (link)].
Milon B., Mitra S., Song Y., Margulies Z., Casserly R., Drake V., Mong J.A., Depireux D.A, & Hertzano R. (2018). The impact of biological sex on the response to noise and otoprotective therapies against acoustic injury in mice. Biology of Sex Differences, 9, 12.
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8

Pharmacological Interventions in Animal Models

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Suberoylanilide hydroxamic acid (SAHA) was obtained from Selleck Chemical (Houston, TX, USA). SAHA was dissolved in DMSO at 50 mg/ml to be delivered in a volume that would bypass the potential detrimental effects of excessive DMSO, as described previously.40 (link) Kanamycin (Sigma, St. Louis, MO, USA) was dissolved in 0.9% saline (Hospira, Lake Forest, IL, USA) and delivered by subcutaneous injection. Furosemide (Hospira) was administered by intraperitoneal injection. Tamoxifen (Sigma) was dissolved in corn oil at 25 mg/ml and delivered by intraperitoneal injection at 0.25 mg/g.
Layman W.S., Williams D.M., Dearman J.A., Sauceda M.A, & Zuo J. (2015). Histone deacetylase inhibition protects hearing against acute ototoxicity by activating the Nf-κB pathway. Cell Death Discovery, 1, 15012-.
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9

Chemical Purity Verification Protocol

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All chemicals and reagents were purchased from Sigma Aldrich (St. Louis, Missouri, USA) unless otherwise stated. MG149 was purchased from Axon Medchem (Groningen, The Netherlands). The purity of MG149 was assessed by HPLC, MS, and NMR by Axon Medchem and was > 99%. Suberoylanilide hydroxamic acid (SAHA) was purchased from Selleckchem (Huissen, The Netherlands). The purity of SAHA was assessed by HPLC, MS, and NMR by Selleckchem and was > 99%.
van den Bosch T., Leus N.G., Wapenaar H., Boichenko A., Hermans J., Bischoff R., Haisma H.J, & Dekker F.J. (2017). A 6-alkylsalicylate histone acetyltransferase inhibitor inhibits histone acetylation and pro-inflammatory gene expression in murine precision-cut lung slices. Pulmonary pharmacology & therapeutics, 44, 88-95.
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10

Desipramine and SAHA Administration in Mice

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Desipramine hydrochloride was purchased from Sigma (St-Louis, MO, USA). Mice received desipramine in the drinking water at a concentration of 100 mg/l, which corresponds to approximately 20 mg/kg of body weight/day. Desipramine solution was changed every week and concentration was adjusted depending on the weight gain and average water consumption of the mice. Suberoylanilide hydroxamic acid (SAHA, also known as vorinostat) was purchased from Selleck Chemicals (Houston, TX, USA). Mice received SAHA through daily intra-peritoneal injections. SAHA was first dissolved in DMSO at a dose of 50 mg/ml. This stock solution was diluted 1:10 every day in saline solution (final concentration: 5 mg/ml) prior to injections at a dose of 25 mg/kg of body weight. Vehicle groups were injected with a solution of 10% DMSO in saline.
Meylan E.M., Halfon O., Magistretti P.J, & Cardinaux J.R. (2016). The HDAC inhibitor SAHA improves depressive-like behavior of CRTC1-deficient mice: possible relevance for treatment-resistant depression. Neuropharmacology, 107, 111-121.
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