Hdac7

Manufactured by Santa Cruz Biotechnology

Sourced in United States

HDAC7 is a histone deacetylase enzyme that catalyzes the removal of acetyl groups from histone proteins, which can lead to changes in chromatin structure and gene expression. The core function of HDAC7 is to regulate gene transcription by modulating the acetylation state of histones.

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Anti-HDAC7

5 protocols using hdac7

Investigating Histone Modifications

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Insulin, TO901317, AA, and TSA were purchased from Sigma Aldrich (St. Louis, MO, USA). Anti-ACC, acetyl-histone H3, acetyl-histone H4, histone H2A, and HDAC5 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-SREBP1, α-Tubulin, HDAC1, HDAC2, HDAC7, and HDAC10 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Shin H.S., Lee Y., Shin M.H., Cho S.I., Zouboulis C.C., Kim M.K., Lee D.H, & Chung J.H. (2021). Histone Deacetylase 1 Reduces Lipogenesis by Suppressing SREBP1 Transcription in Human Sebocyte Cell Line SZ95. International Journal of Molecular Sciences, 22(9), 4477.

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Melanoma Cell Lines Characterization

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The human melanoma cell lines A375 and A2058 used in this study were obtained and grown as previously described [32 (link)]. The following reagents and primary antibodies were used: HDAC1, rabbit, Santa Cruz SC-7872; HDAC2, mouse, Santa Cruz SC-9959; HDAC3, rabbit active motif 40968 IB; HDAC4, rabbit, active motif 40969; HDAC5, mouse, Santa Cruz SC-133225; HDAC6, rabbit, Santa Cruz SC-11420; HDAC7, mouse, Santa Cruz SC-74563; HDAC8, mouse, Santa Cruz SC-17778; anti-HDAC9 antibody (ab18970), Abcam; anti-HDAC10 antibody (ab18971), Abcam; anti-HDAC11, ab18973, abcam; anti-ERK (tERK), anti-phospho-ERK (pERK), anti-AKT (tAKT), anti-phospho-AKT-T308 (pAKT-T308), anti-phospho-AKT-S473 (pAKT-S473), anti-caspase 3, anti-cleaved caspase 3, anti-caspase 8, anti-cleaved caspase 8, anticaspase 9, anti-cleaved caspase 9 (Cell Signaling Technology, Danvers, MA, USA), anti-EGFR (tEGFR), anti-Cathepsin-D, anti-VEGF, anti-MMP9, anti-E-cadherin, and anti-GAPDH (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). Lipofectamine 2000 reagent was obtained from Invitrogen (Carlsbad, CA, USA, Cat. No 11668-019).

Liu J., Gu J., Feng Z., Yang Y., Zhu N., Lu W, & Qi F. (2016). Both HDAC5 and HDAC6 are required for the proliferation and metastasis of melanoma cells. Journal of Translational Medicine, 14, 7.

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Protein Expression and Immunoblotting

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Proteins purified from DNA affinity precipitation assay and the corresponding 1.5% input protein (loading control) as mentioned above were added to 4X Laemmli sample buffer and denatured by boiling for 10 min at 100°C on a heat block. Next, protein samples were resolved on a 4%–20% gradient sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto nitrocellulose membranes. Various selected proteins were detected using mouse anti-GATA1, GATA2, myocyte enhancer factor 2A (MEF2A), MEF2C, CTCF, AT-rich interaction domain 3A (ARID3A), histone deacetylase (HDAC) 2, HDAC4, HDAC5, HDAC7 and TAFII (all purchased from Santa Cruz Biotechnology) and visualised using Super Signal West Dura substrate (Thermo Fisher Scientific) on the Omega Lum C imaging system (Gel Company).

Kong X, & Sawalha A.H. (2019). Takayasu arteritis risk locus in IL6 represses the anti-inflammatory gene GPNMB through chromatin looping and recruiting MEF2–HDAC complex. Annals of the rheumatic diseases, 78(10), 1388-1397.

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Protein Expression Profiling in Kidney Injury

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Antibodies to HDAC4, HDAC5, HDAC7, and HDAC9 were purchased from Santa Cruz Biotechnology (Dallas, TX, United States). Antibodies to NGAL, BMP7, and LC3 were purchased from Abcam Inc (Cambridge, MA United States). Antibodies to p-p53, p53, caspase-3, bcl-2, Bax, PCNA, P-ERK1/2, ERK1/2, E-Cadherin, and β-Actin were purchased from Cell Signaling Technology (Danvers, MA, United States). Antibodies to Atg7 and Beclin-1 were purchased from Arigo Institute (Taiwan, China). Antibodies to GAPDH and α-tubulin were purchased from Proteintech Group Institute (Wuhan, China). Secondary antibodies were purchased from LI-COR Biosciences (Lincoln, NE, United States). Serum creatinine (Scr) reagent kits and Blood urea nitrogen (BUN) reagent kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). All other chemicals were purchased from Sigma (St. Louis, Missouri, United States).

Li J., Yu C., Shen F., Cui B., Liu N, & Zhuang S. (2022). Class IIa histone deacetylase inhibition ameliorates acute kidney injury by suppressing renal tubular cell apoptosis and enhancing autophagy and proliferation. Frontiers in Pharmacology, 13, 946192.

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Evaluating Extracellular Matrix Proteins

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Equal amounts of protein from whole-cell extracts (50 µg/lane) were electrophoresed on 12% sodium dodecylsulfate-polyacrylamide gels, transferred to nitrocellulose membranes, and probed with antibody against HDAC7 (1:100; Santa Cruz Biotechnology), PAI-1 (1:300; Abcam, Cambridge, UK), fibronectin (1:300; Abcam), collagen I (1:300; Abcam), collagen IV (1:300; Abcam), smooth muscle α-actin (1:300; Sigma-Aldrich), or β-actin (1:6,000; Abcam).

Kang D.H., Yin G.N., Choi M.J., Song K.M., Ghatak K., Minh N.N., Kwon M.H., Seong D.H., Ryu J.K, & Suh J.K. (2018). Silencing Histone Deacetylase 7 Alleviates Transforming Growth Factor-β1-Induced Profibrotic Responses in Fibroblasts Derived from Peyronie's Plaque. The World Journal of Men's Health, 36(2), 139-146.

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