Cells used in this study included human osteoblasts (hFOB1.19) and OS cell lines (HOS, U2OS, Saos2, and 143B), all of which were supplied by ATCC (Manassas, VA, USA) and were grown in Dulbecco’s modified Eagle’s medium consisting of 1% penicillin-streptomycin and 10% fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA) in an incubator under an atmosphere of 5% CO2 at 37 C, and HOS and U2OS cells were allocated into the following groups: the si-NC (negative control) group, si-EBLN3P group, si-ANXA3 group, si-HuR group, pc-NC (pcDNA3.1 empty vector) group, pc-ANXA3 group, pc-HuR group, si-HuR + si-NC group, si-HuR + si-EBLN3P group, si-EBLN3P + pc-HuR group, and EBLN3P + pc-ANXA3 group. si-NC, si-EBLN3P, si-ANXA3, si-HuR, pc-NC, pc-ANXA3, and pc-HuR plasmids were designed and completed by GenePharma (Shanghai, China). The target sequences of siRNAs were as follows: si-NC: 5′-UUCUCCGAACGUGUCACGUTT-3′; si-EBLN3P: 5′-TGCAGGGCCAGTGATTGGTTT-3′; si-HuR, 5′-TTGTTAGTGTACAACTCATTT-3′; si-ANXA3, 5′-GATATCTCTCAAGCCTATTAT-3′. Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) was deployed for cell transfection. The concentration of plasmids used for transfection was 800 ng/μL. Further experimentation was carried out after 48 h of cell transfection.
Si nc
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The Si-NC is a lab equipment product manufactured by Thermo Fisher Scientific. It is a silicon-based nanocrystal material used for various research and analytical applications. The core function of the Si-NC is to serve as a versatile nanomaterial with unique optical and electronic properties.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (107 mentions)
Si nc, by GenePharma (71 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (45 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (23 mentions)
Si nc, by RiboBio (21 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (14 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (14 mentions)
Mir nc, by Thermo Fisher Scientific (14 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (13 mentions)
Pcdna3.1 vector, by Thermo Fisher Scientific (11 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (9 mentions)
Si nc, by Sangon (8 mentions)
Sh nc, by Thermo Fisher Scientific (8 mentions)
Mir nc, by GenePharma (7 mentions)
Pcdna nc, by Thermo Fisher Scientific (7 mentions)
Nc mimic, by Thermo Fisher Scientific (6 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (6 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (6 mentions)
Pcdna3, by Thermo Fisher Scientific (6 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (5 mentions)
269 protocols using si nc
1
Osteosarcoma Cell Line Genetic Manipulation
2
ANGPTL2 Knockdown in Glioma Cells
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ANGPTL2-specific siRNA (siANGPTL2) and a control siRNA (siNC) were purchased from Thermo Fisher Scientific. LN18 and T98G cells were cultured in normal medium and grown to 80% confluency before transfection with siANGPTL2 or siNC using Lipofectamine 2000 (Invitrogen). Forty-eight hours after transfection, knockdown efficiency was assessed by Western blot.
3
Modulating miR-206 and c-Met in CRC
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miR-206 mimic, mimic negative control (mimic NC), miR-206 inhibitor, inhibitor NC, c-Met small interfering (si)RNA targeting c-Met (si-c-Met) and si-NC (scramble siRNA) were purchased from Guangzhou RiboBio Co., Ltd. The sequences of miR-206 mimic and inhibitor were as follows: miR-206 mimic, 5′-UGGAAUGUAAGGAAGUGUGUGG-3′; mimic NC, 5′-UUGUACUACACAAAAGUACUG-3′; miR-206 inhibitor, 5′-CCACACACUUCCUUACAUUCCA-3′; and inhibitor NC, 5′-CAGUACUUUUGUGUAGUACAA-3′. The sequences of si-c-Met and si-NC were as follows: si-c-Met sense, 5′-GGUGUUGUCUCAAUAUCAATT-3′, and antisense, 5′-UUGAUAUUGAGACAACACCTT-3′; si-NC sense, 5′-UUCUCCGAACGUGUCACGUTT-3′ and antisense, 5′-ACGUGACACGUUCGGAGAATT-3′. CRC cells were transfected with oligonucleotides using Lipofectamine® 2000 reagent (Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions. Briefly, CRC cells were seeded into 6-well plates at a density of 1×105 cells/ml and transfected with 50 nM oligonucleotides or scrambled duplexes using transfection reagent at 37°C in a humidified incubator with 5% CO2 for 48 h. Transfection efficiency was assessed via reverse transcription-quantitative PCR or western blot analyses 48 h post-transfection.
4
Downregulation of Ecto-5′-nucleotidase in Gastric Cancer
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Small interfering RNA targeting Ecto-5′-nucleotidase (si-Ecto-5′-nucleotidase) and a scrambled negative control (si-NC) were purchased from Thermo Fisher Scientific. Gastric cancer cells were cultured 24 h, and transiently transfected with either si-Ecto-5′-nucleotidase or si-NC by using Lipofectamine® RNAiMAX Transfection Reagent (Invitrogen, Carlsbad, CA, U.S.A.) according to the manufacturer’s instructions. Knockdown efficiency was estimated by Western blot.
5
Regulation of PVT1 and miR-93-5p in Chondrocytes
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Small interfering RNA (siRNA) targeting PVT1 (si-PVT1) and negative control (si-NC) were procured from Shanghai GenePharma Co., Ltd., as were miRNA mimics and inhibitors targeting miR-93-5p (miR-93-5p and anti-miR-93-5p) and their negative control (miR-NC and anti-miR-NC). The sequence of high mobility groupprotein B1 (HMGB1; accession: NM_001363661) was cloned into the pcDNA3.1 vector (pcDNA; Invitrogen; Thermo Fisher Scientific, Inc.) to construct the overexpression vector of HMGB1 (HMGB1). Oligonucleotides (si-PVT1 (40 nM), si-NC (40 nM), miR-93-5p (50 nM), miR-NC (50 nM), anti-miR-93-5p (50 nM), anti-miR-NC (50 nM)) or plasmids were transiently transfected into C28/I2 cells using Lipofectamine® 3000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) and then cultured in DMEM/F-12 containing 5 µg/ml LPS for 48 h. The sequences were: si-PVT1 (5′-GGGUACUGGAAGUAGAAUAUU−3′) and si-NC (5′-UUCUCCGAACGUGUCACGUTT−3′).
6
Hypoxia-Inducible Factor Silencing in BeWo Cells
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Two pairs of HIF1α and HIF2α specific siRNA (si-HIF1α, si-HIF2α) and a non-specific siRNA (si-NC) were obtained from Thermo Fisher Scientific. BeWo cells were seeded on a six-well plate at a density of 30–40% confluence. At 12 hours after seeding, si-HIF1α, si-HIF2α or si-NC were transfected according to manufacturer’s instructions using lipofectamine (Thermo Fisher Scientific) and were further incubated for 24 hours. Culture media were changed and cells were cultured under 20% O2 or 2% O2 condition for additional 24 hours.
7
Silencing TRPM2-AS lncRNA Effects
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LncRNA TRPM2-AS siRNAs (si-TRPM2-AS) and respective negative control sequences (si-NC) were synthesized by Genepharma (Shanghai, China). The sequences were as belows: si-TRPM2-AS sense: 5’-CCACCAGCCACUUACUCAU-3’, si-TRPM2-AS antisense: 5’-AUGAGUAAGUGGCUGGUGG-3’; si-NC sense: 5’-UUCUCCGAACGUGUCACGUTT-3’, si-NC antisense: 5’-ACGUGACACGUUCGGAGAATT-3’.
SiRNAs transfections were performed following the instructions of lipofectamine 3000 regent (Thermofisher, USA). In brief, KYSE-520 and ECA-109 cells were seeded into 6-well plates, respectively. When cell confluence reached 80–90%, lipofectamine 3000 and siRNAs were diluted with opti-MEM (Gibco BRL), respectively. Then, both diluted reagents were mixed and kept at room temperature for 5 min. The lipid-mixture were added on cells. Cells were cultured for additional 48 h followed by subsequent experiments, i.e., CCK-8 assay, transwell assay, flow cytometry, colony formation, and Western blotting.
SiRNAs transfections were performed following the instructions of lipofectamine 3000 regent (Thermofisher, USA). In brief, KYSE-520 and ECA-109 cells were seeded into 6-well plates, respectively. When cell confluence reached 80–90%, lipofectamine 3000 and siRNAs were diluted with opti-MEM (Gibco BRL), respectively. Then, both diluted reagents were mixed and kept at room temperature for 5 min. The lipid-mixture were added on cells. Cells were cultured for additional 48 h followed by subsequent experiments, i.e., CCK-8 assay, transwell assay, flow cytometry, colony formation, and Western blotting.
8
Nrf2 Inhibition in H9c2 Cells
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H9c2 cells were seeded in 6-well plates and grew to about 80% confluence, and then incubated with a serum-free medium. The Nrf2 inhibitor (siNrf2) and negative control (siNC) were designed and synthesized by GenePharma (Shanghai, China). The sequence of Nrf2 siRNA is 5’-GAGGAUGGGAAACCUUACUTT-3′, and the sequence of siNC is 5’-CCAGAATGUAGUGTACGUGAU-3′. H9c2 cells were transfected with siNrf2 and siNC using Lipofectamine 3000 (Invitrogen). The efficiency of inhibition on Nrf2 was verified by western blot analysis at 48 h.
9
Knockdown of SNHG20 in Oral Squamous Cell Carcinoma
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A total of two small interfering RNAs (siRNAs) against SNHG20 were purchased from Shanghai GenePharma Co., Ltd. The siRNAs sequences were as follows: si-NC, 5′-GGATACGGAGTACTATAGC-3′; si-SNHG20-1, 5′-GCCUAGGAUCAUCCAGGUUTT-3′; and si-SNHG20-2, 5′-GCCACUCACAAGAGUGUAUTT-3′. When cell confluence reached 80–90%, 100 nm si-negative control (si-NC), si-SNHG20-1 or si-SNHG20-2 were transfected into SCC4 or SCC15 cells using the Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. Cells were harvested at 48 h after transfection.
10
Lentiviral Transfection of H9c2 Cells
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After the H9c2 cells attained 70–80% confluency, they were transfected with recombinant lentiviruses housing the pHS-AVC-LW1120 (Fendrr) and pHS-BVC-LW280 (negative control) constructs (Syngentech; Beijing, China) for 72 h. Also, the H9c2 cells at 80% confluency were transfected with a nucleolin plasmid (pcDNA3-Nuc) and incubated in a CO2 incubator for 48 h at 37°C. For small interfering RNA (siRNA) transfection, nucleolin or Fendrr siRNA (si-C23 or si-Fendrr) and the corresponding negative control siRNA (si-NC or si-NC; Invitrogen Life Technologies) were transfected into cells. Lipofectamine 2000 reagent (Invitrogen) was used for cell transfection according to the manufacturer's instructions. After 48 h of transfection, the cells were lysed for further experiments.
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