GC cells in the logarithmic growth phase were digested and inoculated onto a 6-well culture plate. After cells reached 60–80% confluence, the miR-335-5p-mimics and miR-335-5p-inhibitor (GenePharma, Shanghai, China) were added to the corresponding wells for further culture for 24–48 h.
Mir 335 5p mimics
Manufactured by GenePharma
Sourced in China
MiR-335-5p mimics are synthetic RNA molecules designed to mimic the function of the natural microRNA MiR-335-5p. MiR-335-5p is involved in the regulation of gene expression. The mimics are used in research applications to study the biological functions of MiR-335-5p.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Mir 335 5p inhibitor, by GenePharma (2 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Puromycin, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Sw480, by American Type Culture Collection (1 mentions)
Small interfering rnas sirna, by GenePharma (1 mentions)
Hieff trans liposomal transfection reagent, by Yeasen (1 mentions)
Pcdna zeb1 as1, by GenePharma (1 mentions)
Jetprime reagent, by Thermo Fisher Scientific (1 mentions)
Rock1 sirna si rock1, by GenePharma (1 mentions)
4 protocols using mir 335 5p mimics
1
GC Cells Transfection with miR-335-5p
2
Bladder Cancer Cell Lines and Transfection Protocols
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Human BC cell lines (EJ, T24, and RT4) and human epithelial SV40 immortalized uroepithelium cell line SV-HUC-1 were obtained from the Stem Cell Bank, Chinese Academy of Sciences in Shanghai, China. Cells were cultured in DMEM medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS (Thermo Fisher Scientific) in 5% CO2 at 37°C.
For transfection, siRNA, miR-335-5p mimics, miR- 335-5p inhibitors, and negative controls were synthesized by GenePharma Co., Ltd (Shanghai, China) and transfected using the Lipofectamine 2000 transfection reagent (Thermo Fisher Scientific) according to the manufacturer’s protocol. For the construction of stable cell line, BC cells were infected with lentiviral-CIRBP-shRNA and lentiviral-control-shRNA (LV-NC) for 24 hours and selected using 5 µg/mL puromycin (Sigma-Aldrich Co., St Louis, MO, USA) for 7 days.
For transfection, siRNA, miR-335-5p mimics, miR- 335-5p inhibitors, and negative controls were synthesized by GenePharma Co., Ltd (Shanghai, China) and transfected using the Lipofectamine 2000 transfection reagent (Thermo Fisher Scientific) according to the manufacturer’s protocol. For the construction of stable cell line, BC cells were infected with lentiviral-CIRBP-shRNA and lentiviral-control-shRNA (LV-NC) for 24 hours and selected using 5 µg/mL puromycin (Sigma-Aldrich Co., St Louis, MO, USA) for 7 days.
3
Modulating ZEB1-AS1 and APOC1 in CRC
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The human CRC cell lines HCT116 and SW480, purchased from the American Type Culture Collection (ATCC), were cultured in RPMI-1640 medium (Gibco, MD, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin/streptomycin (Gibco; Thermo Fisher Scientific, Inc.) in a humidified atmosphere of 5% CO2 at 37° C.
Small interfering RNAs (siRNAs) specifically targeting ZEB1-AS1 or APOC1, negative control siRNA (si-NC), miR-335-5p mimics and miR-335-5p inhibitor were all obtained from GenePharma Corporation (Shanghai, China). A plasmid vector, pcDNA-ZEB1-AS1 or pcDNA-APOC1, which could consistently express ZEB1-AS1 or APOC1, was constructed by GenePharma Corporation. Cell transfection of siRNA, miRNA mimic or inhibitor and plasmid vectors were all conducted with Hieff Trans™ Liposomal Transfection Reagent (Yeason, Shanghai, China) and Opti-MEM (Gibco) for 48h, according to the manufacturer’s protocols.
Small interfering RNAs (siRNAs) specifically targeting ZEB1-AS1 or APOC1, negative control siRNA (si-NC), miR-335-5p mimics and miR-335-5p inhibitor were all obtained from GenePharma Corporation (Shanghai, China). A plasmid vector, pcDNA-ZEB1-AS1 or pcDNA-APOC1, which could consistently express ZEB1-AS1 or APOC1, was constructed by GenePharma Corporation. Cell transfection of siRNA, miRNA mimic or inhibitor and plasmid vectors were all conducted with Hieff Trans™ Liposomal Transfection Reagent (Yeason, Shanghai, China) and Opti-MEM (Gibco) for 48h, according to the manufacturer’s protocols.
4
Transfection of miR-335-5p and ROCK1 siRNA
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A549 and SPC-A1 cells were seeded in 6-well plates. When cells had reached 40–60% confluence, we performed transfection in accordance with the manufacturer’s instructions using jetprime reagent (Invitrogen). Cells were collected at 48–72 h after transfection for further experiments. MiR-335-5p mimics, ROCK1 siRNA (Si-ROCK1) and corresponding control were purchased from GenePharma company (Suzhou, China). The target siRNA sequences were as follows: ROCK1si-1 5′-GCAGAUGAAACAGGAAAUA-3′, ROCK1 si-2 5′-GGCAGAGGAAGAAUAUAAA-3′.
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