Ezh2 sirna

Manufactured by Santa Cruz Biotechnology

Sourced in United States

EZH2 siRNA is a small interfering RNA (siRNA) designed to target the EZH2 gene. EZH2 is a histone methyltransferase that plays a role in regulating gene expression. The EZH2 siRNA can be used to study the function of the EZH2 gene in cellular processes.

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4 protocols using ezh2 sirna

Molecular Mechanism of 3-DZNeP and Cisplatin

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3-DZNeP was purchased from Selleckchem (Houston, TX, USA). Cisplatin was purchased from Sigma (St. Louis, MO, USA). Antibodies to caspase3, E-cadherin, EZH2, H3K27me3, p-JNK, p-ERK1/2, p-p38, histone H3, and tubulin were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibody to NGAL was purchased from R&D systems (Minneapolis, MN, USA). Antibodies to GAPDH, β-actin, horseradish peroxidase-conjugated secondary antibodies, enhanced chemiluminescence (ECL) detection kit, cell counting kit-8 (CCK-8), and DAPI were from Beyotime institute of Biotechnology (Jiangsu, China). Scr and BUN reagent kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). EZH2 siRNA, E-cadherin siRNA and scramble siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Ni J., Hou X., Wang X., Shi Y., Xu L., Zheng X., Liu N., Qiu A, & Zhuang S. (2019). 3-deazaneplanocin A protects against cisplatin-induced renal tubular cell apoptosis and acute kidney injury by restoration of E-cadherin expression. Cell Death & Disease, 10(5), 355.

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EZH2 Inhibition and Modulation in SSc

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DZNep (Cayman Chemicals), an EZH2 inhibitor, was dosed at 0.2–5 μM for fibroblasts and 5 μM for ECs for 48 h, with PBS as a negative control. When indicated, another EZH2 inhibitor, GSK126 (Cayman Chemicals), was dosed at 0.5–10 μM for 72 h in SSc dermal fibroblasts. Cell viability was checked using Trypan blue and was not affected by either EZH2 inhibitor with the dosages used. To evaluate the effect of EZH2 on angiogenesis in ECs, we used 75 nM EZH2 siRNA (Santa Cruz Biotechnology) to transfect ECs for 48 h. Overexpression of EZH2 in ECs was achieved by transfecting 0.33 μg of EZH2 (control vector pCMV6-XL5; Origene) using Lipofectamine 2000 (Invitrogen) in T12.5 flasks. After 5 h, culture media were changed to EGM supplemented with bovine brain extract (Lonza). Matrigel tube formation assay was performed 24 h after transfection. Overexpression of EZH2 was also done in fibroblasts, using 0.1 μg of either control or EZH2 vector in a 12-well plate for 24–72 h. Successful transfection was confirmed by qPCR.

Tsou P.S., Campbell P., Amin M.A., Coit P., Miller S., Fox D.A., Khanna D, & Sawalha A.H. (2019). Inhibition of EZH2 prevents fibrosis and restores normal angiogenesis in scleroderma. Proceedings of the National Academy of Sciences of the United States of America, 116(9), 3695-3702.

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EZH2 Silencing in ESCC Cells

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To inhibit EZH2, 50 nmol/L EZH2 siRNA (Santa cruze) was transfected into ESCC cells using Lipofectamine 2000 reagent according to the manufacturer's instructions. Cells transfected by the transfection agent with no siRNA (Mock) and with scramble‐controlled siRNA (Negative control) were employed as controls. The cells were harvested 48 hours after transfection.

Ding X., Qi C., Min J., Xu Z., Huang K, & Tang H. (2020). Long non‐coding RNA HEIH suppresses the expression of TP53 through enhancer of zeste homolog 2 in oesophageal squamous cell carcinoma. Journal of Cellular and Molecular Medicine, 24(18), 10551-10559.

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Silencing EZH2 with siRNA

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The EZH2 siRNA was obtained from Santa Cruz Biotechnology; miR-138-5p mimic, inhibitor, and negative control were synthesized by Guangzhou RiboBio. The transfection we had used Lipofectamine 2000 (Invitrogen) according to the corresponding's protocol.
EZH2 siRNA
5′-TTCGAGCTCCTCTGAATCAAA-3′

Zeng T., Luo L., Huang Y., Ye X, & Lin J. (2021). Upregulation of miR-138 Increases Sensitivity to Cisplatin in Hepatocellular Carcinoma by Regulating EZH2. BioMed Research International, 2021, 6665918.

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