Small interfering RNAs (siRNAs) targeting Serotonin-receptor 7 (5-HT7) and Forkhead box M1 (FOXM1) genes and non-silencing control siRNA were purchased from Sigma-Aldrich. Exponentially growing cells were plated 24h before transfection and transfected with 5-HT7 siRNA, two different FOXM1 siRNAs or control siRNA at a nal concentration of 50 nM and 100 nM for 72h, using HiPerFect Transfection Reagent (Qiagen, Valencia, CA) according to the manufacturer's protocol. non-silencing control siRNA-transfected cells were used as negative controls. After treatment, the cells were harvested and processed for further analysis [21, 25, (link)26 ].
Control non silencing sirna
Manufactured by Merck Group
Sourced in United States
Control non-silencing siRNA is a laboratory reagent used in RNA interference (RNAi) experiments. It is a small interfering RNA (siRNA) that does not target any known gene, serving as a negative control for RNAi experiments. This product is used to assess the effects of siRNA transfection without the influence of target gene knockdown.
Automatically generated - may contain errors
Lab products found in correlation
5 ht7 sirna, by Qiagen (2 mentions)
Hiperfect transfection reagent, by Qiagen (2 mentions)
Ru486, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Taqman probe, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
Progesterone, by Merck Group (1 mentions)
Charcoal stripped fbs, by Thermo Fisher Scientific (1 mentions)
Calcein am fluorescent dye, by BD (1 mentions)
Growth factor reduced matrigel, by BD (1 mentions)
Medroxyprogesterone acetate mpa, by Merck Group (1 mentions)
Opti mem serum free media, by Thermo Fisher Scientific (1 mentions)
X tremegene transfection reagent, by Merck Group (1 mentions)
Egfr sirna, by Thermo Fisher Scientific (1 mentions)
5 protocols using control non silencing sirna
1
Silencing 5-HT7 and FOXM1 Genes
2
Silencing 5-HT7 and FOXM1 Genes
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Small interfering RNAs (siRNAs) targeting Serotonin-receptor 7 (5-HT7) and Forkhead box M1 (FOXM1) genes and non-silencing control siRNA were purchased from Sigma-Aldrich. Exponentially growing cells were plated 24h before transfection and transfected with 5-HT7 siRNA, two different FOXM1 siRNAs or control siRNA at a nal concentration of 50 nM and 100 nM for 72h, using HiPerFect Transfection Reagent (Qiagen, Valencia, CA) according to the manufacturer's protocol. non-silencing control siRNA-transfected cells were used as negative controls. After treatment, the cells were harvested and processed for further analysis [21, 25, (link)26 ].
3
Breast Cancer Cell Line Signaling
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Dulbecco's modified Eagles medium (DMEM) and phenol red-free DMEM, glutamine, penicillin, streptomycin, Fetal Bovine Serum (FBS), charcoal stripped FBS and TaqMan probes were purchased (Life Technologies, Carlsbad, CA). 17β-estradiol (E2), R5020, RU486, progesterone and medroxyprogesterone acetate (MPA) were purchased from Sigma Aldrich (Saint Louis, MO). Growth factor reduced matrigel (Cat# 356231) and calcein AM fluorescent dye (Cat# 354216) were purchased from BD Biosciences (San Jose, CA). PR-B directed siRNAs [60 (link), 61 (link)] and control non-silencing siRNA (Cat# SIC001) were ordered from Sigma Aldrich (St. Louis, MO). siRNAs targeting TM4SF1 (Cat# S8367), HES1 (Cat# S6920), PRKCH (CAT#S1107), and ELF5 (CAT# S4629) were purchase from Life Technologies (Carlsbad, CA).
4
Cloning and Luciferase Assay for MALAT1
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The malat1 fragment M1‐M5(NR_002819.2) was cloned into the pcDNA vector (Figure 1 A). pGMSP1‐Luc (SP1 luciferase reporter plasmid containing SP1‐responsive element) was from Yeasen Company (China). Validated duplex siRNAs for malat1 were purchased from Sigma Chemical Co. (St Louis, MO, USA).15 Control non‐silencing siRNA also was purchased from Sigma.
5
Targeted Knockdown of Key Proteins in TNBC
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For siRNA-mediated knockdown, control non-silencing siRNA (Sigma-Aldrich, catalog no. SIC001), EGFR siRNA (Thermo Fisher Scientific, catalog no. AM51331), ITGA5 siRNA (Sigma-Aldrich, catalog no. SASI_Hs01_00058581), and FAK siRNA (SMARTpool, L-003164-00) were used. A total of 2.0 × 105 MDA-MB-231-4175 TNBC cells were plated in a 6-well plate 24 hours before transfection. On the day of transfection, cells were approximately 50% confluent. siRNA transfections were performed by combining 10 mL Xtreme-gene transfection reagent (Sigma-Aldrich, SITRAN-RO) diluted in 100 mL Opti-MEM serum-free media (Gibco, 31-985-070), with 10–20 nmol/L siRNA diluted in 100 mL Opti-MEM and incubated for 20 minutes at 15°C–25°C. The complex was added to 2 mL of medium and the mixture was then added to the plated cells for 24–48 hours before each experiment, depending on the specific gene of interest. Gene knockdown was validated using Western blotting.
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