Normal human liver L02 cells and the HCC cell lines (Huh-6, Huh-7, SNU-449 and Li-7) were purchased from the Institute of Biochemistry and Cell Biology of the Chinese Academy of Sciences (Shanghai, China). All the cells were cultured in DMEM medium (Gibco, Grand Island, NY, U.S.A.) containing 10% fetal bovine serum (Gibco). Cell transfection was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, U.S.A.) according to the manufacturer’s instructions. Negative control inhibitor (NC inhibitor), miR-183-5p inhibitor, miR-183-5p mimics and small interfering RNA specifically targeting PDCD4 (si-PDCD4) were synthesized by GenePharma Co. Ltd (Shanghai, China). When cells were allowed to grow to 70–90% confluence, cell transfection was performed. NC inhibitor or miR-183-5p inhibitor at a final concentration of 50 nM was transfected into HCC cells. For the rescue assay, si-PDCD4 at a final concentration of 50 nM was transfected into miR-183-5p inhibitor-treated HCC cells. The sequence information of NC inhibitor, miR-183-5p inhibitor, si-PDCD4, negative control mimics and miR-183-5p mimics was listed as follows: NC inhibitor, 5′-UUCUCCGAUCUGGCUACAGU-3′; miR-183-5p inhibitor, 5′-UUCUGACCAUCUUAAGUGAU-3′; si-PDCD4, 5′-GGAGGUGGAUGUGAAAGAU-3′; negative control mimics, 5′-ACUACUGAGUGACAGUAGA-3′; miR-183-5p mimics, 5′-UAUGGCACUGGUAGAAUUCACU-′3.
Mir 183 5p mimics
Manufactured by GenePharma
The MiR-183-5p mimics is a laboratory reagent designed to mimic the function of the miR-183-5p microRNA. MicroRNAs are small non-coding RNA molecules that play a role in gene expression regulation. This product is intended for use in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Mir 183 5p inhibitor, by GenePharma (1 mentions)
Si pdcd4, by GenePharma (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Mir nc, by GenePharma (1 mentions)
2 protocols using mir 183 5p mimics
1
Regulation of PDCD4 by miR-183-5p in HCC
2
Osteosarcoma Cell Transfection Optimization
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MiR-183-5p mimics, miR-183-5p inhibitor and their corresponding negative controls (miR-NCs) were obtained from GenePharma (Shanghai, China). Osteosarcoma cells were seeded in a 6-well plate (3×105 per well) and cultured to 70‑80% density. Next, the cell transfection process was performed using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.). Lipofectamine 2000 (Thermo Fisher Scientific, Inc.) was prepared in a sterile Eppendorf tube, and 5 µL of Lipofectamine 2000 and 100 µL of serum‑free medium were incubated at room temperature for 5 min. siRNA (50 nmol) and 100 µL of serum‑free medium were incubated at room temperature for 20 min. The cells in the culture flask were washed. Serum-free medium (without antibiotics) was added to the complex, which was then mixed, and the mixture was added into the 50 mL culture flask for transfection. The flask was placed in an incubator containing 5% CO2 at 37°C for 6–8 h, and the reagent was then replaced with complete culture medium. Further experiments were conducted following 48 h after transfection.
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