After cells were plated onto 6-well plates for western blotting and then 96-well plates for cell viability assays for 24 h, they were transiently transfected using the Lipofectamine RNAiMAX reagent (catalog no. 13778; Invitrogen, Carlsbad, CA) with 20 nM SMARTpool Human KRas siRNA (catalog no. M-005069-00; Dharmacon), SMARTpool NT siRNA #2 (catalog no. D-001206-14-05; Dharmacon), SignalSilence® GSK3α/β siRNA (catalog no. 6301; Cell Signaling), and SignalSilence® control siRNA (unconjugated; catalog no. 6568; Cell Signaling). Transfected cells were collected 48 or 72 h after transfection for western blot analysis as described above, and after 72 h for CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, WI, USA) as described below.
Signalsilence control sirna
Manufactured by Cell Signaling Technology
Sourced in United States
SignalSilence® Control siRNA is a laboratory tool used for RNA interference (RNAi) experiments. It is a non-targeting control siRNA designed to have minimal sequence homology to known genes. The product is intended to serve as a negative control in RNAi studies.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (7 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (5 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions)
Signalsilence stat3 sirna 2, by Cell Signaling Technology (3 mentions)
Signalsilence gsk3α β sirna, by Cell Signaling Technology (2 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (2 mentions)
Hiperfect transfection reagent, by Qiagen (2 mentions)
Celltiter glo luminescent cell viability assay, by Promega (1 mentions)
Signalsilence p38 mapk sirna, by Cell Signaling Technology (1 mentions)
Akt sirna, by Cell Signaling Technology (1 mentions)
P44 42 mapk erk1 2 sirna, by Cell Signaling Technology (1 mentions)
Amaxa nucleofector technology, by Lonza (1 mentions)
Bamhi, by New England Biolabs (1 mentions)
Anti actin, by Merck Group (1 mentions)
Sb415286, by Merck Group (1 mentions)
Adeasy xl adenoviral vector system, by Agilent Technologies (1 mentions)
Recombinant gsk 3β, by New England Biolabs (1 mentions)
Genjet plus transfection reagent, by SignaGen (1 mentions)
Lithium chloride (licl), by Merck Group (1 mentions)
Dual luciferase reporter dlr assay system, by Promega (1 mentions)
28 protocols using signalsilence control sirna
1
Transfection and Cell Viability Assay
2
Investigating p38 MAPK Signaling in U87MG Cells
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U87MG cells were seeded at 100000 cells per well into six-well plates and grown for 24 h. Transfection of siRNA (50 nM) was performed using Lipofectamine 2000 (Thermo Fisher Scientific). SignalSilence p38 MAPK siRNA (#6564) and SignalSilence control siRNA (#6568) from “Cell Signaling Technology” were used. Control siRNA (Fluorescein conjugate) (#6201) was added to both preparations (at ratio 1:5). 48 h after transfection, cells were treated by CBD for 6 h (for Western blot analysis) or 24 h for cell-cycle apoptosis analysis using PI staining of fixed cells and the subsequent FACS analysis of green transfected cells.
3
Silencing Rab43, ERK1/2, and Akt in PANC1 Cells
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After a 2-day culture, PANC1 cells were reseeded into six-well plates at 2 × 105 cells/well. After 24 h, the Rab43 siRNA (50 pmol, Santa Cruz Biotechnology Inc., Dallas, TX, USA), p44/42 MAPK (ERK1/2) siRNA (100 nM) and Akt siRNA (100 nM, Cell Signaling Technology) were delivered to cells via a lipid-mediated Lipofectamine 2000 Transfection Reagent. A nonrelated scramble siRNA (Invitrogen) and SignalSilence control siRNA (Cell Signaling Technology) were used as a transfection control.
4
Akt1/Akt2 Knockdown in Rat β-Cells
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Knockdown of Akt1 and/or Akt2 in rat primary β-cells was achieved by transfection with specific siRNA (Akt1: SASI_Rn01-00063656, Akt2: SASI_Rn01_00047688, Sigma–Aldrich; and control siRNA (Signal Silence Control siRNA, Cell Signaling Technology, Danvers, MA). In brief, liposome-siRNA, Lipofectamine 2000 (Life Technologies, Inc.) and 100 nm siRNA was prepared in 200 μl of opti–modified Eagle's medium as described by the suppliers. Transfected primary β-cells were incubated 72 h to allow siRNA expression before cell treatment and analysis.
5
β-Catenin Silencing in Breast Cancer Cells
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Briefly, T47D and MCF-7 cells (5×104/well) were collected and seeded in a 6-well plate. Once the cells reached 95% confluence, they were trans-fected with a SignalSilence® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions. The final concentration of siRNA was 100 nmol/l. The sequences of siRNA were as follows: β-catenin siRNA forward, 5′-UGG UUG CCU UGC UCA ACA A-3′ and reverse, 5′-ACC AAC GGA ACG AGU UGU U-3′; and control siRNA forward, 5′-CGG UUA ACC UGC UAG AU-3′ and reverse, 5′-UGG CAU ACG GUA UCU AG-3′. At 24 h post-transfection, the cells were collected for subsequent analyses.
6
Cofilin-1 Knockdown in hUCM-MSCs
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To knockdown Cofilin-1, hUCM-MSCs were transfected at P2 according to the manufacturer’s instructions on TCPS or custom-made 40:1 PDMS using lipofectamine 3000 (Thermo Fisher Scientific) with 150 nM of SignalSilence® Cofilin siRNA I or SignalSilence® Control siRNA as control (Cell Signalling Technology). 5-FUrd incorporation experiments were performed 72 h after transfection.
7
Investigating MKK7 Regulation of Cell Proliferation
8
Transfection and Adenoviral Transduction in MODE-K Cells
9
Comprehensive Reagent and Resource List
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Cycloheximide, actinomycin D, MG132, insulin, LiCl, SB415286 and DAPI were purchased from Sigma-Aldrich. The restriction endonucleases (NheI, XhoI, XbaI, EcoRI, KpnI and BamHI) and recombinant GSK-3β were bought from New England Biolabs. All of the primers used were synthesized from Integrated DNA Technologies. Protein A/G plus agarose was purchased from Santa Cruz. The anti-HA, anti-Myc antibodies and SignalSilence Control siRNA, SignalSilence GSK-3α/β siRNA were obtained from Cell Signaling; anti-actin was obtained from Sigma-Aldrich; anti-SCFFbw7 was obtained from Abcam and anti-SREBP-1 was bought from Becton-Dickinson and Co. GenJet Plus transfection reagent was purchased from SignaGen Laboratories and Halt™ combined protease and phosphatase inhibitor cocktails were bought from Thermo Scientific. Trypsin and Lys-C enzymes for mass spectrometry and the Dual-Luciferase® Reporter (DLR™) Assay System were obtained from Promega. SCFFbw7 siRNA 1 (s30664), SCFFbw7 siRNA 2 (s224357), SimplyBlue and Lipofectamine RNAiMAX were purchased from Invitrogen. AdEasy XL Adenoviral Vector System was purchased from Agilent Technologies.
10
Cell Culture and Treatment Protocol
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MBAMB231, SKBR3, and U87 cells were maintained in RPMI (Roswell Park Memorial Institute) medium containing 10% fetal bovine serum, while NIH-3T3 cells were grown in DMEM (Dulbecco’s Modified Eagle’s Medium) containing 10% calf serum. Signal Silence Control siRNA (Cell Signaling, Catalog No. 6568) and Signal Silence Survivin siRNA II (Cell Signaling, Catalog No. 6546) were introduced into cells with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). Cells were treated with 50 nM PTX (Sigma, St. Louis, MO, USA), 1 µM nocodazole (Sigma), 5 µM PD98059 (Cell Signaling, Danvers, MA, USA), 10 µM BPTES (EMD Millipore, Darmstadt, Germany), 10 µM 968 (EMD Millipore), 2 µM doxorubicin (Sigma), 5 µM fluorouracil (Sigma), 5 µM etoposide (Sigma), 2 µM cisplatin (Sigma), 2 µM Gefitinib (LC Laboratories, Woburn, MA, USA), 5 µM AG538 (EMD Millipore), 1 µM cytochalasin D (Sigma), and 5 µM 17-AAG (Sigma).
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