The H9c2 cardiomyocytes were seeded in 30,000 cells per square cm in six-well plates or 96-well plates, and transfection began at a cell density of 30%-50% by using the commercially available siRNA transfection reagent (RIBOBIO, Cat. No. C10511-05). H9c2 cells were transfected with 100 nM of Smpd1small interfering RNA (RIBOBIO, siRNA-Smpd1, 5′-GCTACCGAGTTTACCAAAT-3′) or negative control (NC, 5′-UUCUCCGAACGUGUCACGUTT-3′) according to the manufacturer’s protocol. After 24 h transfection, cells were treated with specific concentrations of drugs, followed by a total incubation for another 24 h.
Sirna transfection reagent
Manufactured by RiboBio
Sourced in China
The SiRNA transfection reagent is a laboratory product designed to facilitate the delivery of small interfering RNA (siRNA) molecules into cells. It is a tool used in molecular biology and genetic research to study gene expression and function.
Automatically generated - may contain errors
Lab products found in correlation
Scrambled sirna, by RiboBio (3 mentions)
Hdac3 sirna duplex, by RiboBio (2 mentions)
Sirna duplex, by RiboBio (2 mentions)
Hdac2, by RiboBio (1 mentions)
Hdac1, by RiboBio (1 mentions)
Rabbit anti lpl polyclonal antibody sc 32885, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti β actin polyclonal antibody sc 130656, by Santa Cruz Biotechnology (1 mentions)
Simplechip enzymatic chromatin ip kit, by Cell Signaling Technology (1 mentions)
Aβ25 35, by Bachem (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Rgfp966, by Selleck Chemicals (1 mentions)
8 protocols using sirna transfection reagent
1
Transfection and Treatment of H9c2 Cells
2
Knockdown of P2X7 Receptor in Cell Lines
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Cells were transfected with 20 nM of siRNA for 8 h with siRNA transfection reagent (RiboBio, Guangzhou, China) to knockdown purinergic 2X7 receptor (P2X7R). Briefly, cells were treated with 100 nM of PMA for 48 h and washed in fresh medium without antibiotics. Afterward, the cells were treated with siRNA duplex solution for 8 h. The medium was subsequently replaced with normal culture medium. Control cells were transfected with scrambled sequence siRNA control (RiboBio, Guangzhou, China). The cell lysates were utilized for Western blot analysis to verify the efficacy of protein knockdown by siRNA.
3
Targeting HDAC1-3 Modulates p75NTR and Histone Acetylation
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HDAC1, HDAC2, and HDAC3 siRNA duplex (Guangzhou RIBOBIO CO, LTD) were used to interfere with endogenous HDAC1, HDAC2 and HDAC3 mRNA levels, respectively. Cells were seeded in six-well culture microplates at a density of 1 × 105 cells/well in 2 ml of antibiotic-free normal growth medium. The following day, cells were transfected with 50 nM siRNA duplex and 12 μl of siRNA transfection reagent (Guangzhou RIBOBIO CO, LTD) mixed in DMEM/F12 (1:1) media with 1% fetal bovine serum. The transfected cells were incubated at 37°C for 24 h. Untreated cells and non-specific siRNA (scrambled siRNA; Guangzhou RIBOBIO CO, LTD) were used as controls. The interference efficiency was evaluated by qRT-PCR and western blot analyses. In addition, the expression levels of p75NTR mRNA and protein, and Ace-H3K9 and Ace-H4K12 levels were investigated by the above methods. This experiment was performed in duplicate and repeated three times.
4
Aβ25-35 Peptide Aggregation and ChIP Assay
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Aβ25–35, a toxic fragment of the full-length Aβ peptide, was purchased from American Peptide Company (131602-53-4, United States), which needs to be solubilized in sterile water and aggregated at 37°C for 7 days before use. The siRNA transfection reagent was purchased from Guangzhou RiboBio Company Limited (C10511-1, Guangzhou, China). The Simple ChIP Enzymatic Chromatin IP Kit was obtained from Cell Signaling Technology (9003S, BOS, USA), and rabbit anti-acetylation of histone 3 lysine 9 (Ace-H3K9) polyclonal antibody for ChIP was from Merck Millipore (06942, Darmstadt, Germany). The rabbit anti-PPARγ polyclonal antibody (D262458), rabbit anti-HDAC2 polyclonal antibody (D155199), and rabbit anti-HDAC3 polyclonal antibody (D260481) were purchased from Bio Basic Inc. (Canada); rabbit anti-LPL polyclonal antibody (sc-32885) and rabbit anti-β-actin polyclonal antibody (sc-130656) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Goat anti-rabbit immunoglobulin G (lgG) secondary antibody was from Shanghai Sangon Biotech Company Limited (D111018, Shanghai, China). Cell culture medium was purchased from American Hyclone Inc. (SH30023.01B, USA).
5
Modulating CDK1, STAT3, and HDAC3 in Pancreatic Cancer
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EP300-core, Flag-HDAC3, CDK1-WT-myc, CDK1-K33R-myc, CDK1-K33Q-myc, STAT3-WT-myc were constructed in pEGFP-N3-Vectors. The plasmids were transfected into cells using Lipofectamine 3000 (Invitrogen, USA). CDK1, STAT3 and HDAC3 siRNAs (Ribobio, China) were used to knock down CDK1, STAT3 and HDAC3 gene expression. The CDK1, STAT3, HDAC3 and control siRNA (100 pM) were transfected into PANC-1, HPC-Y5 cells using siRNA transfection reagent (Ribobio, China) 24h before fisetin treatment. The cells were collected after fisetin treatment for 48h. The shRNA of CDK1 (5ʹ-GTGGAATCTTTACAGGACTAT-3ʹ) was constructed in pLKO.1 vector for stable down-regulating CDK1 in PANC-1 cells. Infected cell populations were selected using 2 mg/mL puromycin.
6
Silencing HDAC1, 2, and 3 in SH-SY5Y cells
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HDAC1, 2, and 3 small-interfering RNA (siRNA) duplexes (Guangzhou RiboBio Co, Ltd, China) were used simultaneously (si-HDAC1-2-3) or singly (si-HDACs) to interfere with endogenous HDAC1, HDAC2, and HDAC3 expression. The siRNA oligos were as follows: HDAC1: 5′-CCGGTCATGTCCAAAGTAA-3′; HDAC2: 5′-TCCGTAATGTTGCTCGATG-3′; and HDAC3: 5′-GCATTGATGACCAGAGTTA-3′. In brief, differentiated SH-SY5Y cells were seeded in 6-well culture microplates at a density of 1 × 105 cells/well in 2 mL of antibiotic-free normal growth medium. The following day, cells were transfected with a solution of 50 nmol/L siRNA duplex and 12 μL of siRNA transfection reagent (Guangzhou RiboBio Co, Ltd) mixed in DMEM/F12 medium containing 10% FBS. The transfected cells were incubated at 37°C for 24 hours. The untreated (non-siRNA) and nonspecific siRNA treated (scrambled siRNA; Guangzhou RiboBio Co, Ltd) cells were used as controls. The interference efficiency was evaluated by qRT-PCR and Western blot analyses. In addition, the expression levels of TrkA mRNA and protein were investigated by the above methods. This experiment was performed in duplicate and repeated 3 times.
7
TLR4 Knockdown in BV2 Cells
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BV2 cells were transfected with 100 nM of siRNA for 8 h with siRNA transfection reagent (RiboBio, Guangzhou, Guangdong, China) to knockdown the expression of TLR4. Briefly, cells were treated with siRNA duplex solution for 12 h after washing with fresh medium without antibiotics. The medium was subsequently replaced with culture medium. Control cells were transfected with scrambled siRNA sequence (RiboBio, Guangzhou, China). Cell lysates were utilized for western blotting to verify the efficacy of protein knockdown.
8
HDAC3 Modulation Regulates AQP4 and miR-130a
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HDAC3 siRNA duplex (Guangzhou RiboBio Co., Ltd.) or RGFP966 (Selleck Chemicals Co., Ltd.) was used to interfere with endogenous HDAC3 mRNA levels. siRNA was performed with siRNA transfection reagent (Guangzhou RiboBio Co., Ltd.) as we have described in detail previously (Zhang et al., 2017 (link)). Cells were incubated in 6-well culture microplates at 37°C with antibiotic-free medium containing 10 μM RGFP966. After 24 h, the expression of AQP4 mRNA and protein, and miR-130a levels were investigated by the above methods. Untreated cells and non-specific siRNA (scrambled siRNA; Guangzhou RiboBio Co., Ltd.) were used as controls. This experiment was repeated three times and performed in duplicate.
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