Synthetic, chemically modified short single- or double-stranded RNA oligonucleotides: miR-26a mimics, miR-26a control (Ctrl), anti-miR-26a, anti-miR-26a Ctrl, p68 siRNA (sip68), p53 siRNA (sip53), and EZH2 siRNA (siEZH2) were synthesized from Shanghai GenePharma Co., Ltd. Commercial miR-26a expression Lentivirus and counterpart control vector were purchased from Shanghai GenePharma Co., Ltd. Commercial p68 expression plasmid and sh.p53 plasmid carrying small hairpin RNA targeting p53 were purchased from Shanghai Gene-Chem Co., Ltd. Commercial mutp53 expression plasmids and wild-type p53 expression plasmid were constructed and provided by GuangZhou Biosicen Biotechnology CO., Ltd. All oligonucleotide sequences are listed in supplementary table 1 .
Ezh2 sirna siezh2
Manufactured by GenePharma
Sourced in China
EZH2 siRNA (siEZH2) is a laboratory reagent designed to target and silence the expression of the EZH2 gene. It is a synthetic, double-stranded RNA molecule that can be used to study the functional role of EZH2 in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Mir 26a mimic, by GenePharma (1 mentions)
Anti mir 26a, by GenePharma (1 mentions)
Pcdna vector, by GenePharma (1 mentions)
Mimics nc, by Thermo Fisher Scientific (1 mentions)
Si ezh2, by Thermo Fisher Scientific (1 mentions)
Si scramble, by GenePharma (1 mentions)
Mimics nc, by GenePharma (1 mentions)
Dznep, by Merck Group (1 mentions)
Nc oligonucleotides, by GenePharma (1 mentions)
3 protocols using ezh2 sirna siezh2
1
Synthetic RNA Oligonucleotides for Cellular Manipulation
2
Regulating EZH2 Expression in Oral Cancer Cells
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The miR-144-3p mimics and mimics negative control (mimics NC) were obtained from GenePharma. The EZH2 overexpression vector, pcDNA-EZH2, and pcDNA vector were constructed by GenePharma. In addition, EZH2 siRNA (si-EZH2) and si-Scramble were also purchased from GenePharma.
CAL-27 and SCC-4 cells (8.0×105/well) in a 6-well plate grown to approximately 80% confluence, and then respectively transfected with miR-144-3p mimics (20 nM), mimics NC (20 nM), si-EZH2 (50 nM), or 2 µg pcDNA-EZH2 using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.). In addition, inhibition experiments were performed using the EZH2 inhibitor, GSK126 (a compound competes with S-adenosyl-methionine for binding to EZH2, thereby inhibiting histone methyltransferase activity without affecting EZH2 protein expression), as previously described (16 (link)). Briefly, the CAL-27 and SCC-4 cells were treated with 5 µM GSK126 (Shanghai HanXiang Life Technology Limited Corporation) or DMSO for 48 h and then collected for use in further experiments. The concentration of DMSO used was ≤1% to ensure the lack of cytotoxicity.
CAL-27 and SCC-4 cells (8.0×105/well) in a 6-well plate grown to approximately 80% confluence, and then respectively transfected with miR-144-3p mimics (20 nM), mimics NC (20 nM), si-EZH2 (50 nM), or 2 µg pcDNA-EZH2 using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.). In addition, inhibition experiments were performed using the EZH2 inhibitor, GSK126 (a compound competes with S-adenosyl-methionine for binding to EZH2, thereby inhibiting histone methyltransferase activity without affecting EZH2 protein expression), as previously described (16 (link)). Briefly, the CAL-27 and SCC-4 cells were treated with 5 µM GSK126 (Shanghai HanXiang Life Technology Limited Corporation) or DMSO for 48 h and then collected for use in further experiments. The concentration of DMSO used was ≤1% to ensure the lack of cytotoxicity.
3
Modulating EZH2 in Glioblastoma Cells
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EZH2 siRNA (siEZH2) and negative control (NC) oligonucleotides were purchased from GenePharma (Shanghai, China, see Additional file 1 ). Oligonucleotides were transected into U87, U251, and GL261 cells using the transfection reagent Lipofectamine2000 (Invitrogen) following the manufacturer’s instructions with oligonucleotides at a working concentration of 100 nM. The transfected cells were incubated for 24 or 48 h before subsequent experiments.
The EZH2 functional inhibitor, DZNep, was purchased from Sigma-Aldrich. DZNep was added to U87 and U251 cell medium at a concentration of 5 μM for indicated time duration.
The EZH2 functional inhibitor, DZNep, was purchased from Sigma-Aldrich. DZNep was added to U87 and U251 cell medium at a concentration of 5 μM for indicated time duration.
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