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Hdac6

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HDAC6 is a histone deacetylase enzyme that plays a role in the regulation of cellular processes. It is involved in the deacetylation of various proteins, including histones, which can influence gene expression and other cellular functions. The core function of HDAC6 is to remove acetyl groups from target proteins, thereby modulating their activity and localization.

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Lab products found in correlation

High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Hdac3, by Cell Signaling Technology (1 mentions) Hdac4, by Thermo Fisher Scientific (1 mentions) Hdac5, by Cell Signaling Technology (1 mentions) Total h3, by Cell Signaling Technology (1 mentions) Acetylated h3k9 k14, by Cell Signaling Technology (1 mentions) Hdac1, by Cell Signaling Technology (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Bcl2 associated x (bax), by Thermo Fisher Scientific (1 mentions) Hdac1, by Thermo Fisher Scientific (1 mentions) Hdac2, by Thermo Fisher Scientific (1 mentions) Erk1 and erk2 sirna, by Santa Cruz Biotechnology (1 mentions) Nucleofector kit 5, by Lonza (1 mentions) On targetplus smartpool sirna, by Thermo Fisher Scientific (1 mentions) Cell line nucleofector kit 5, by Lonza (1 mentions) Mda mb 231, by American Type Culture Collection (1 mentions) Bt549, by American Type Culture Collection (1 mentions) Hek293t, by American Type Culture Collection (1 mentions) Gapdh, by Thermo Fisher Scientific (1 mentions)

6 protocols using hdac6

1

Histone Acetylation and HDAC Profiling

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Histone preparations were made from young and aged mice BMSCs. Equal amounts of core histones were resolved by 15% SDS-PAGE, transferred onto PVDF membranes and probed with the following antibodies: acetylated H3K9/K14 (#9677, Cell Signaling Technology, Danvers, MA, USA), acetylated H4K12 (#13944, Cell Signaling Technology, Danvers, MA, USA), HDAC1 (#34589, Cell Signaling Technology, Danvers, MA, USA), HDAC3 (#85057, Cell Signaling Technology, Danvers, MA, USA), HDAC4 (MA5-15580, ThermoFisher scientific), HDAC5 (#98329, Cell Signaling Technology, Danvers, MA, USA), HDAC6 (PA1-41056, ThermoFisher scientific), total H3 (#4499, Cell Signaling Technology, Danvers, MA, USA), total H4 (#13919, Cell Signaling Technology, Danvers, MA, USA) and β-actin (#4970, Cell Signaling Technology, Danvers, MA, USA).
Ma C., Gao J., Liang J., Dai W., Wang Z., Xia M., Chen T., Huang S., Na J., Xu L., Feng S., Dai K, & Liu G. (2021). HDAC6 inactivates Runx2 promoter to block osteogenesis of bone marrow stromal cells in age-related bone loss of mice. Stem Cell Research & Therapy, 12, 484.
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2

Silencing Key Regulators in MM Cells

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Short interfering (si)RNA for BIM, BAX, BAK, HDAC1, HDAC2 and HDAC6 were purchased from Thermo Fisher Scientific. ERK1 and ERK2 siRNA were purchased from Santa Cruz Biotechnology. siRNA were electroporated into MM cell lines using the Lonza nucleofector kit V (Lonza, Basel, Switzerland). The manufacturer’s G-15 program was used for KMS18 and OPM2; O-23 was used for MM1S and KMS28. All experiments were performed in triplicate.
Ramakrishnan V.G., Miller K.C., Macon E.P., Kimlinger T.K., Haug J., Kumar S., Gonsalves W.I., Rajkumar S.V, & Kumar S.K. (2019). Histone deacetylase inhibition in combination with MEK or BCL-2 inhibition in multiple myeloma. Haematologica, 104(10), 2061-2074.
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3

siRNA Transfection Protocol Using Amaxa

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For small interfering RNA (siRNA) transfection, scrambled, HDAC6 and cereblon (CRBN) ‘ON-TARGETplus SMARTpool siRNA' were purchased from Thermo Scientific (Lafayette, CO, USA). siRNA transfection was carried out by Amaxa electroporation system using ‘Cell Line Nucleofector Kit V' solution, according to manufacturer's protocol (Lonza, Koln, Germany).
Hideshima T., Cottini F., Ohguchi H., Jakubikova J., Gorgun G., Mimura N., Tai Y.T., Munshi N.C., Richardson P.G, & Anderson K.C. (2015). Rational combination treatment with histone deacetylase inhibitors and immunomodulatory drugs in multiple myeloma. Blood Cancer Journal, 5(5), e312-.
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4

Characterization of NEDD9 and Cortactin Mutants

Cited 1 time
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Authenticated cell lines MDA-MB-231, BT549, HEK293T were purchased from American Type Culture Collection, MDA-231-LN (Caliper Life Sci.) and grown based on the manufacturer's recommendations. NEDD9 wild type and knockout fibroblasts, vectors expressing human full-length NEDD9 or truncation mutants of NEDD9 were previously described (30 (link)). The shRNA/siRNAs expressing constructs against NEDD9, HDAC6, AURKA, CTTN and the control were purchased from ThermoFisher Scientific (sequences available upon request). Mouse wild type, C- and N-term CTTN truncation mutant constructs were previously described (31 (link)). The acetylation-null 9KR cortactin construct was generated by site-directed mutagenesis using full-length mouse cortactin. Tissue culture medium and supplements were purchased from ATCC. Cells were transfected with plasmid DNA and/or siRNA using nucleofection (Amaxa).
Kozyreva V.K., McLaughlin S.L., Livengood R.H., Calkins R.A., Kelley L.C., Rajulapati A., Ice R.J., Smolkin M.B., Weed S.A, & Pugacheva E.N. (2014). “NEDD9 Regulates Actin Dynamics through Cortactin Deacetylation in an AURKA/HDAC6-dependent Manner”. Molecular cancer research : MCR, 12(5), 681-693.
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5

Gene Expression Analysis of Adipose-Derived Stem Cells

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Total RNAs of ASCs were extracted with RNeasy Mini kit (QIAGEN, Hilden). Reverse transcription was performed using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Darmstadt), as instructed. All probes for gene analysis were obtained from Applied Biosystems: ADIPOQ (#Hs00605917_m1), AURKA (#Hs01582072_m1), AURKB (#Hs00945858_g1), CCP110 (#Hs00206922_m1), PLK1 (#Hs00153444_m1), PLK4 (#Hs00179514_m1), KIF2A (#Hs00189636_m1), KIF24 (#Hs00950248_m1), HDAC6 (#Hs00195869_m1), SMO (#Hs01090242_m1), GLI1 (#Hs00171790_m1), NANOG (#Hs04260366_g1), PTCH1 (#Hs00181117_m1), RUNX2 (#Hs01047973_m1), KLF4 (#Hs00358836_m1), c-MYC (#Hs00153408_m1), KLF6 (#Hs00810569_m1), PPARγ (#Hs01115513_m1), LEPTIN (#Hs00174877_m1), OCT4 (#Hs04260367_gH), SOX2 (#Hs01053049_s1), and GAPDH (#Hs02758991_g1). Real-time PCR was performed with a StepOnePlus Real-time PCR System (Applied Biosystems). The data were analyzed using StepOne Software v.2.3 (Applied Biosystems) as described previously [11 (link)].
Ritter A., Kreis N.N., Roth S., Friemel A., Jennewein L., Eichbaum C., Solbach C., Louwen F, & Yuan J. (2019). Restoration of primary cilia in obese adipose-derived mesenchymal stem cells by inhibiting Aurora A or extracellular signal-regulated kinase. Stem Cell Research & Therapy, 10, 255.
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6

Quantifying miRNA and mRNA Expression

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Total RNA was purified with miR-Neasy Mini Kit (Qiagen, Valencia, CA, USA), cDNA was transcribed from 10ng RNA using TaqMan® miRNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA). Real-time PCR was performed using TaqMan® miRNA assays (Applied Biosystems, Foster City, CA, USA) for miR-22 (#000398) according to the manufacturer's instructions. Expression of U6 (#001973) was used as a reference. For quantification of expression of pri-miR-22 or various mRNAs, RNA extraction was performed with the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). cDNA was subsequently transcribed using the High Capacity cDNA Reverse Transcription Kit followed by PCR analysis using TaqMan® Gene Expression Assays: pri-miR-22 (#Hs03302632_pri), NCOA1 (#Hs00186661), CDK6 (Hs 01026371_m1), HDAC4 (Hs 01041638_m1), HDAC6 (Hs 00195869_m1), MAX (Hs 00811069_g1), MYCBP (Hs 00429315_g1), PTEN (Hs 02621230_s1) (all from Applied Biosystems, Foster City, CA, USA) according to the manufacturer's instructions. GAPDH (#Hs02758991_g1) was included as a reference. The amplification was performed on an Mx3005P qPCR System (Agilent Technologies, Santa Clara, CA, USA) real-time cycler on standard settings.
Sibbesen N.A., Kopp K.L., Litvinov I.V., Jønson L., Willerslev-Olsen A., Fredholm S., Petersen D.L., Nastasi C., Krejsgaard T., Lindahl L.M., Gniadecki R., Mongan N.P., Sasseville D., Wasik M.A., Iversen L., Bonefeld C.M., Geisler C., Woetmann A, & Odum N. (2015). Jak3, STAT3, and STAT5 inhibit expression of miR-22, a novel tumor suppressor microRNA, in cutaneous T-Cell lymphoma. Oncotarget, 6(24), 20555-20569.
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