All the overexpression plasmids targeting VHL (pcDNA3.1-VHL), HIF-1α (pcDNA3.1-HIF-1α), and ZEB1 (pcDNA3.1-ZEB1) were designed and synthesized by GeneChem (China), and the empty plasmid was used as a negative control. HIF-1α siRNA (siHIF-1α), VHL siRNA (siVHL), ELK1 siRNA (siELK1), ZEB1 siRNA (siZEB1), and corresponding negative control were purchased from RiboBio (Guangzhou, China). MiR-21-5p mimics, miR-21-5p inhibitors, and matched control (mimic-NC or inhibitor-NC) were synthesized by Invitrogen (Shanghai, China). All the small interference RNAs were transfected at a final concentration of 50 nM, and the plasmid was transfected at a final concentration of 0.2 μg for 96 well plates and 1.6 μg for 12 well plates. Lipofectamine 3000 (Invitrogen, USA) was used for cell transfections according to the manufacturer’s instructions. Protein and total RNA were extracted after 48 h.
Mir 21 5p mimic
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The MiR-21-5p mimic is a synthetic RNA molecule designed to mimic the function of the naturally occurring microRNA miR-21-5p. MicroRNAs are small, non-coding RNA molecules that play a role in regulating gene expression. The MiR-21-5p mimic can be used in research applications to study the biological functions and regulatory mechanisms of miR-21-5p.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (5 mentions)
Mir 21 5p mimic, by GenePharma (3 mentions)
Inhibitor nc, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Pcdna3, by Thermo Fisher Scientific (2 mentions)
Nc mimic, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Si meg 3, by Thermo Fisher Scientific (1 mentions)
Si meg3, by GenePharma (1 mentions)
Sh hnrnpa2b1, by GenePharma (1 mentions)
Mir 21 5p mimic, by RiboBio (1 mentions)
Pcdna3.1 vector, by GenePharma (1 mentions)
Si nc, by GenePharma (1 mentions)
Mir nc mimic, by Thermo Fisher Scientific (1 mentions)
Pcdna3, by Addgene (1 mentions)
Mir 21 5p mimic, by Sangon (1 mentions)
Mimic nc, by Sangon (1 mentions)
7 protocols using mir 21 5p mimic
1
Overexpression and Knockdown of Key Regulators
2
Regulation of NSCLC by HNRNPA2B1
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HNRNPA2B1 overexpression plasmids, lentivirus-mediated HNRNPA2B1 shRNA (sh-HNRNPA2B1, 5ʹ-CCGTAAGCTCTTTATTGGTGGCTTA-3ʹ), si-MEG3, miR-21-5p mimics and inhibitor were purchased from GenePharma (Shanghai, China). The sh-NC, vector and miR-NC were used as the control groups. NSCLC cell lines were planted in 6-well plates 24 h prior to sh-HNRNPA2B1, si-MEG3, miR-21-5p mimics or inhibitor transfection with 50–60% confluence, and then mixed with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacture instructions.
3
miR-21-5p Mimic Transfection Protocol
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The miR-21-5p mimic and its negative control (NC) were synthesized by RiboBio (Guangzhou, China). Wild-type cells (5.0×105 cells/well) were grown in 6-well plates in 2 ml culture medium. When cell confluence reached 50%–60%, the miR-21-5p mimic and its negative control were treated with Lipofectamine 2000 reagent (Invitrogen, USA) according to the manufacturer’s instructions. The cells were transfected in Opti-MEM (Gibco, USA) for 8 h, and then the medium was changed to normal culture medium. After 48 h, the cells were harvested for western blot and qRT-PCR. All groups were plated in 6-well culture plates at the same time and were harvested 48 h later.
4
Overexpressing CASC7 in Breast Cancer
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The lentivirus system (oe-CASC7 and oe-NC, MOI = 50) was used to infect the breast cancer cells to stably overexpression CASC7. Compared with the overexpression of whole full-length CASC7 lentiviral production (oe-CASC7), oe-NC is an empty lentiviral vector used as a control. Both oe-CASC7 and oe-NC lentiviral vector production are obtained from Genechem, Shanghai, China. Oe-CASC7 and oe-NC breast cancer cells were collected after incubating with 1 mg/ml puromycin for 72 h. About at 70% confluence, cells were transfected with miR-21-5p mimic (100 pmol) and corresponding negative control (100 pmol, Invitrogen, Shanghai, China) by Lipofectamine™2000 (ThermoFisher, USA) to conduct follow-up experiments.
5
Transfection of miR-21-5p and CDKN2C in Melanoma
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The miR‐21‐5p mimic, inhibitor and their negative controls (NCs) were purchased from GenePharma Co. Ltd. (Shanghai, China). Small interference RNA for CDKN2C (si‐CDKN2C) and its NC (si‐NC) were also synthesized by GenePharma Co. Ltd. The CDKN2C overexpression plasmid was constructed by inserting CDKN2C cDNA into a pcDNA3.1 vector by GenePharma Co. Ltd. For transfection, A375 or M14 cells were seeded at a density of 2 × 105 cells per well in a six‐well culture dish, and transfection was performed when 80% confluence was achieved. A total of two 8‐µL (500 ng·µL−1) plasmids (pcDNA3.1‐CDKN2C, si‐CDKN2C) or mimics (miR‐21‐5p mimic, inhibitor and NC) and 8 µL Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) were suspended in 100 µL Opti‐MEM (Gibco), and the final concentration of CDKN2C plasmids or mimics used was 1 mg·mL−1. Then, the mixture was added into the cell culture and incubated for 48 h.
6
Regulation of MAPK10 by miR-21-5p
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miR-21-5p mimic and negative control (NC) were synthesized by Shanghai GenePharma Co., Ltd. pcDNA3-MAPK10 and pcDNA3 were purchased from Addgene (cat. no. 13758). A549 (1×103/well) and H1299 (1×103/well) cells were seeded in 96-well plates, cultured for 24 h and transfected with 50 nM miR-21-5p mimic, 50 nM miR-NC mimic, 2 µg pcDNA3 or 2 µg pcDNA3-MAPK10 using Lipofectamine 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. After 48 h, A549 and H1299 cells were collected for subsequent experiments.
7
Modulating miR-21-5p in Aβ-Induced hNSCs
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StemProÒ human neural stem cells (hNSCs) (H9-derived) were purchased from ThermoFisher (USA). The medium complete was used for the optimal growth and expansion of hNSCs and to maintain the undifferentiated state of hNSCs as previously described (Mcc et al. 2021 ). The cells were divided into four groups, blank control, Aβ group with 5 μM Aβ1-42 (cat: P2000022, PLlabs, Canada) treatment for 24 h, AuNPs (Yaphank, USA) group with 10 ppm AuNPs treatment for 24 h, 10 ppm AuNPs were added into Aβ group for another 24 h. hNSCs were transfected with miR-21-5p inhibitor, inhibitor-NC, miR-21-5p mimic and mimic-NC using Lipofectamine 2000 (cat: no. 11668019, Invitrogen, USA). inhibitor-NC, inhibitor, miR-21-5p mimic and mimic-NC were purchased from Sangon Biotech (Shanghai). The sequence of NC and inhibitor were as follows: miR-21-5p inhibitor, 5'-UCAACAUCAGUCUGAUAAGCUA-3'; inhibitor negative control, 5'-CAGUACUUUUGUGUAGUACAAA-3'; miR-21-5p mimic, 5'-AACAUCAGUCUGAUAAGCUAUU-3'; mimic-NC, 5'-UUGUACUACACAAAAGUACUG-3'.
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