siRNA targeting PRL-3 (D-006859-04), HSP60 (D-010600-02) and Src (D-003175-05) were purchased from ThermoFisher Scientific. Negative knockdown control siRNA (4390843) was from Life Technologies. For siRNA transfection, 2 × 105 cells were transfected with 60 pmol of siRNA molecules using lipofectamine RNAiMAX (Life Technologies) according to the manufacturer’s instructions.
Sirna targeting
Manufactured by Thermo Fisher Scientific
SiRNA targeting is a laboratory tool used for gene silencing. It involves the use of small interfering RNA (siRNA) molecules to target and suppress the expression of specific genes within cells. The core function of this product is to enable researchers to study gene function and explore potential therapeutic applications.
Automatically generated - may contain errors
Lab products found in correlation
Ici 182 780, by Bio-Techne (2 mentions)
Sineg, by Thermo Fisher Scientific (2 mentions)
Phlda1 miscript target protector for mir 181, by Qiagen (2 mentions)
Mir 181a and b synthetic mimics, by Qiagen (2 mentions)
Anti mir 181a and b inhibitors, by Thermo Fisher Scientific (2 mentions)
4 oht, by Merck Group (2 mentions)
β actin antibody, by Merck Group (2 mentions)
Il 1β, by R&D Systems (2 mentions)
Tnf α, by R&D Systems (2 mentions)
Interleukin 6 (il 6), by R&D Systems (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Sybr green pcr master mix, by Roche (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Amaxa nucleofection technology, by Lonza (1 mentions)
Sirna targeting, by Horizon Discovery (1 mentions)
8 protocols using sirna targeting
1
Targeted Knockdown of PRL-3, HSP60, and Src
2
KRAS and PART1 Modulation in Cell Lines
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shRNA targeting KRAS was obtained from Dharmacon and was transduced in NCI-H2444 and NCI-H647 cells. Cells were selected with puromycin (2 µg/mL) for 2 weeks. siRNA targeting PART1 (Assay ID: n260057; 5ʹ- GGAACAACACAGAUGAGAUtt – 3ʹ) or a negative control siRNA (#4390843) were obtained from ThermoFisher Scientific. PART1 overexpression plasmid or negative control vector was obtained from GeneChem (Shanghai, China). For transfection cells were plated in antibiotic-free media. Cells were transfected with control or PART1 siRNA (10 nM final concentration) using Polyplus jetPrime transfection reagent. For transduction, BEAS-2B cells were transduced with either lentiviral particles containing control or PART1 overexpression construct using polybrene. Cells were selected with puromycin (2 µg/mL) for 2 weeks. All indicated treatments were done 48 hours following siRNA transfection. Successful knockdown or overexpression was verified by quantitative real-time polymerase chain reaction (qRT-PCR).
3
Quantifying Gene Expression via RT-qPCR
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Total RNA was extracted using Trizaol (Invitrogen) according to the manufacturer's instructions. Two micrograms of total RNA was subjected to a random-primed reverse transcription using SuperScritpt 2 reverse transcriptase (Invitrogen). Real-time PCR was conducted in triplicates using Applied Biosystem 7900 HT with 5 ng of cDNA, 1 μM of each primer pair and SYBR Green PCR master mix (Roche). The primer sequences were listed in Table S1 . Relative mRNA levels were normalized to GAPDH. SiRNA targeting PR was purchased from Thermo Fisher (Cat N. J-003433-08-0005).
4
Molecular Regulation of PHLDA1 Expression
5
Molecular Regulation of PHLDA1 Expression
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E2, 4OHT, and Act D were purchased from Sigma. TNFα, IL-1β, and IL6 were obtained from R&D Systems. ICI 182,780 (ICI) was purchased from Tocris. The NFκB inhibitor, IKK7, was purchased from EMD Millipore. The PHLDA1 antibody was purchased from Santa Cruz Biotechnology (sc-23866) and the β-actin antibody from Sigma (A5441). DMA was generously provided by Dr. Gregory Thatcher (UIC). siRNA targeting ERα or PHLDA1 or a nonspecific control (siNeg) was purchased from Ambion. PHLDA1 miScript target protector for miR-181 and miR-181a and b synthetic mimics were purchased from Qiagen. Anti-miR-181a and b inhibitors were purchased from Ambion.
6
Silencing p62 via siRNA
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siRNA targeting p62 (s16960 or s16961) and a non-targeting control sequence were obtained from Ambion. Cells were transfected with 20 nM siRNA using Lipofectamine 2000 (Grand Island, NY, Invitrogen) according to manufacturer’s recommendations. p62 knockdown was achieved after 24 h of transfection.
7
HSP27 Silencing Enhances CDDP Sensitivity
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HSC2 cells were transfected with siRNA targeting HSP27 or negative control siRNA (Ambion, Austin, TX). Human HSP27‐specific siRNAs were purchased from Ambion: siRNA #1 sense, 5′‐GCGUGUCCCUGGAUGUCAAtt‐3′ and antisense, 5′‐UUGACAUCCAGGGACACGCgc‐3′; siRNA #2 sense, 5′‐GCCGCCAAGUAAAGCCUUAtt‐3′ and antisense, 5′‐UAAGGCUUUACUUGGCGGCag‐3′. At 48 h after transfection, the cells were incubated in culture medium with or without 10 μM CDDP for an additional 24 h before analysis.
8
Bcl-XL and NIK Knockdown in CLL Cells
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CLL cells were transfected using the Amaxa nucleofection technology (Amaxa) as previously described10 (link) with 3 µg siRNA targeting Bcl-XL and non-targeting siRNA (Ambion, Thermo Fisher Scientific) or 3 µg siRNA targeting NIK and non-targeting siRNA (Dharmacon). After nucleofection, cells were cultured on 3T40L for 24 h before drugs sensitivity assay was performed or protein lysates were obtained.
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