Chinese Academy of Sciences (Shanghai, China) was the provider of the five MM cell lines (MM.1S, U266, RPMI-8226, KM3 and H929) and normal plasma cells (nPCs). Cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific, MA, USA) with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 mg/mL streptomycin (Gibco, Carlsbad, CA, USA) at 37 °C in 5% CO2 with 100% humidity. miR-214-3p mimics, miR-214-3p inhibitors, small hairpin RNA (shRNA) targeting LINC00665 (sh-LINC00665), LINC00665 overexpressing plasmid (pcDNA-LINC00665) and negative controls (miR-NC, in-NC, sh-NC and NC-vector) were obtained from Ribobio (Guangzhou, China). U266, H929 and MM.1S cells were inoculated in a 6-well cell plate with a density of 1 × 105/mL, cultured at 37 °C in 5% CO2 for 16 h, and then transfected. Lipofectamine 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) was adopted to transfect miR-214-3p mimics, miR-214-3p inhibitors, LINC00665 shRNA or pcDNA-LINC00665 into the cells in accordance to the supplier’s instructions.
In nc
Manufactured by RiboBio
Sourced in China
The In-NC is a laboratory instrument used for nucleic acid detection and quantification. Its core function is to perform real-time quantitative PCR (qPCR) analysis, a widely used technique for sensitive and precise measurement of nucleic acid targets.
Automatically generated - may contain errors
Lab products found in correlation
Mir nc, by RiboBio (4 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Sh nc, by RiboBio (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Igf 1, by Thermo Fisher Scientific (1 mentions)
Si nc, by GenePharma (1 mentions)
Mir 301a 3p mimic, by RiboBio (1 mentions)
Mimic nc, by RiboBio (1 mentions)
5 protocols using in nc
1
Regulation of Multiple Myeloma by miR-214-3p and LINC00665
2
Targeted knockdown of RC3H2 using siRNA
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The siRNAs against RC3H2 (si-RC3H2), si-Scramble, or miR-101-3p mimics (miR-101-3p), miR-101-3p inhibitors (in-miR-101-3p), NC mimics (miR-NC) and NC inhibitor (in-NC) (Ribobio, China) were transfected into cells using Lipofectamine RNAiMAX (Invitrogen, USA) at a final concentration of 100 nM. At 24 h after incubating, the cells were harvested and we performed the corresponding experiments. The siRNAs used in our study were designed and synthesized by Guangzhou RiboBio (Guangzhou, China).
3
HUVEC Transfection and Gene Expression
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HUVECs in the logarithmic growth phase were digested, passed, and inoculated into 6-well plates with a density of 5 × 106/well. The cells were transfected when they achieved steady growth. Ribobio Biotechnology Co., Ltd (Guangzhou) took on the design and synthesis of miR-301a-3p mimics, inhibitors, and corresponding negative controls (miR-NC, in-NC). IGF1 overexpression plasmids (IGF1) and the negative control (NC) were purchased from GenePharma (Shanghai, China). IGF1-overexpression plasmids or NC were transfected into HUVECs using Lipofectamine® 3000 (Invitrogen; ThermoFisherScientific, Inc.) as instructed by the supplier. The cells were cultured in an incubator with 5% CO2 at 37°C. Following 48 hours’ transfection, the original medium was replaced by a fresh and complete one for another 24 hours’ culture. With total RNA extracted from the cells, quantitative real-time polymerase chain reaction (qRT-PCR) was implemented to check alterations in the profile of each molecule in the transfected cells [27 (link)].
4
Investigating Circular RNA PITX1 and ITGA6 in NSCLC
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H1975 and A549 cells in the logarithmic growth phase were inoculated onto 60-mm culture plates at 1 × 107 cells/well and incubated at 37°C with 5% CO2 for 24 hours prior to transfection. GenePharma (Shanghai, China) was responsible for the design of the siRNA specifically targeting circ-PITX1 (si-circ-PITX1), siRNA specifically targeting ITGA6 (si-ITGA6), circ-PITX1 overexpression plasmids, and circ-PITX1 negative controls (vector or si-NC). miR-30e-5p mimics, miR-30e-5p inhibitors, and their negative controls (miR-NC and NC-in) were supplied by RiboBio (Guangzhou, China). Lipofectamine 3000 (Thermo Fisher Scientific, IL, USA) was adopted to transfect the above expressing vectors into NSCLC cells in line with the supplier’s instructions. The transfection validity was examined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) 48 hours following the transfection.
5
Modulating miR-301a-3p Expression
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miR-301a-3p mimic (5′-CAGUGCAAUAGUAUUGUCAAAGC-3′) and negative control (NC mimic, 5′-UCUACUCUUUCUAGGAGGUUGUGA-3′) were purchased from Guangzhou RiboBio Co., Ltd. miR-301a-3p inhibitor (5′-GCUUUGACAATCTATTGCACTG-3′) and negative control (NC-in, 5′-ACCGCUAAUCAUACGAAUACAC-3′) were purchased from Guangzhou RiboBio Co., Ltd. Plasmids (pcDNA3.1) expressing BTG1 and lentiviruses expressing miR-301a-3p were purchased from Hanbio Biotechnology Co., Ltd. Oligonucleotide (100 nM) transfection was performed using Lipofectamine® 2000 (Thermo Fisher Scientific, Inc.). After 48 h of transfection, cells were collected for further investigation.
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