Pre-miR-5701 expression vectors and control vector were chemically synthesized in the pcDNA6.2-GW/EmGFP vector (Invitrogen, Carlsbad, CA, USA). The overexpressed plasmid of FGFR2, miR-5701 inhibitor, and all siRNA (i.e., siDNMT3A, siHDAC3, and siMBD1) were purchased from the GenePharma Corporation (SGC, Shanghai, China). The FGFR2 wild-type and mutated mRNA were cloned in between the SacI and XhoI sites of the pmirGLO expression vector (Invitrogen, Carlsbad, CA, USA). All related sequences used are listed in the Supplementary Table 2 . The SGC-7901 cells were treated with 5-Aza (2.5, 5, 7.5, 10, and 15 μM; MedChemExpress, Shanghai, China) for 3 days and TSA (100, 300, 500, 700, and 900 nM; MedChemExpress, Shanghai, China) for 24 h. Then, the total RNA was extracted using the TRIzol protocol.
Si dnmt3a
Manufactured by GenePharma
Sourced in China
Si-DNMT3A is a laboratory reagent used for gene expression analysis. It is a small interfering RNA (siRNA) that targets the DNMT3A gene, which encodes a DNA methyltransferase enzyme involved in DNA methylation. Si-DNMT3A can be used to study the role of DNMT3A in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Si dnmt3b, by GenePharma (3 mentions)
Si dnmt1, by GenePharma (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Pcdna6.2 gw emgfp vector, by Thermo Fisher Scientific (1 mentions)
5 aza, by MedChemExpress (1 mentions)
Sh malat1, by GenePharma (1 mentions)
Oe dnmt1, by GenePharma (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Mir 29a, by GenePharma (1 mentions)
Lipofectamine 2000 kit, by GenePharma (1 mentions)
Nc sirna, by GenePharma (1 mentions)
Oe nc, by Thermo Fisher Scientific (1 mentions)
Pcdna3.1 vector, by Thermo Fisher Scientific (1 mentions)
Si nc, by GenePharma (1 mentions)
Si sp1, by GenePharma (1 mentions)
5 protocols using si dnmt3a
1
Epigenetic Regulation of FGFR2 Expression
2
Molecular Regulation of DNMT and MALAT1
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The overexpression plasmid of DNMT1 (oe-DNMT1), the overexpression plasmid of MALAT1 (oe-MALAT1), the overexpression plasmid of DNMT1 (oe-DNMT1), the small interfering RNA of DNMT1 (si-DNMT1), the small interfering RNA of DNMT3A (si-DNMT3A), the small interfering RNA of DNMT3B (si-DNMT3B), the short hairpin RNA of MALAT1 (sh-MALAT1), and mimics/inhibitor of miR-137 and their negative controls were all obtained from GenePharma Co., Ltd (Shanghai, China), and transfected into cells with Lipofectamine™ 3000 (Invitrogen, CA, USA).
3
Transfection of miR-29a and DNMT3A/B in Cervical Cancer Cells
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miR-29a mimics (miR-29a) and scrambled miRNA (Scrambled), siRNA for DNMT3A (si-DNMT3A), siRNA for DNMT3B (si-DNMT3B) and negative control (siRNA-NC) were purchased from Shanghai GenePharma Co., Ltd., and the sequences are presented in Table I . HeLa and C-33A cells were seeded into 6-well plates at a density of 2×105 cells/well and cultured at 37°C for 24 h, prior to transfection using the Lipofectamine® 2000 kit (cat. no. 11668027; Invitrogen, Thermo Fisher Scientific, Inc.) at 37°C for 48 h, according to the manufacturer's protocol. The final concentrations of miR-29a mimics and siRNAs were 50 nM and 80 nM, respectively. Cells were harvested 24 h post-transfection.
4
Epigenetic Regulation of RARB Expression
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The cells were transfected using Lipofectamine 2000 (11 668 030, Invitrogen Inc., Carlsbad, CA, USA) as per the manufacturer’s instructions. The siRNA targeting DNMT1 mRNA (si-DNMT1), DNMT3A mRNA (si-DNMT3A), DNMT3B mRNA (si-DNMT3B), RARB mRNA (si-RARB) and negative control (si-NC) were acquired from Shanghai GenePharma Co., Ltd. (Shanghai, China). The sequences of the siRNAs are shown in Table 1 . The full-length RARB cDNA was subcloned into pcDNA3.1 vector (Invitrogen) for the construction of RARB overexpression (oe-RARB), with the empty vector as a negative control (oe-NC). After seeding and plating in 6-well plates for 1 d, the cells were transfected with the plasmids. After 48 h of transfection, the cells were stored for later use. Transfection efficiency was determined by RT-qPCR.
To suppress the expression of DNMT1, the cells were treated with 10 μM 5-aza-2′-deoxycitidine (5-AzaC) (189 825, Sigma-Aldrich Chemical Company, St Louis, MO, USA) 3 days before transfection. An equal volume of dimethylsulfoxide (DMSO) was applied as a control.
To suppress the expression of DNMT1, the cells were treated with 10 μM 5-aza-2′-deoxycitidine (5-AzaC) (189 825, Sigma-Aldrich Chemical Company, St Louis, MO, USA) 3 days before transfection. An equal volume of dimethylsulfoxide (DMSO) was applied as a control.
5
Macrophage Induction and Genetic Manipulation
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Macrophages were induced from human monocyte leukemia cell line (THP‐1) as described previously (Guo et al., 2020 (link)). Lv‐miR‐195‐3p, Lv‐miR‐neg, adenovirus encoding IL‐31 (Ad‐IL‐31), adenovirus expressing Sp1 (Ad‐Sp1), Ad‐GFP, siRNAs against Sp1 (si‐Sp1), siRNAs against DNMT3a (si‐DNMT3a), siRNAs against HDAC11 (si‐HDAC11) or control siRNA (si‐NC) were obtained from Genepharma (Shanghai, China) and infected as previously described (Guo et al., 2020 (link)). And they were transiently transfected into the cells using Lipofectamine 2000 (Life Technologies, Gaithersburg, MD, USA) following the manufacturer's instruction. The transfection efficiency was detected by qRT‐PCR and western blot, then the cells were collected for downstream analysis.
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