MiR-103a-3p mimics (5’-AGCAGCAUUGUACAGGGCUAUGA-3’), negative control miRNA (miR-NC: 5’-AUAGCCCUGUACAAUGCUGCUUU-3’), small interfering RNA targeting HMGB1 (siHMGB1: 5’-CUCACCAAGUCUCCUCAAU-3’) and siRNA negative control (siNC: 5’-AUUGAGGAGACUUGGUGAG-3’) were synthesized and purified by GenePharma Co., Ltd (Shanghai, China). These oligonucleotides were cloned into the lentivirus expression vector of Hu6-MCS-CMV-EGFP (GenePharma Co., Ltd). The recombined vector was triple transfected into 80% confluent HEK293T cells with packaging vectors pHelper 1.0 and 2.0 (GeneChem Co., Ltd.) with Lipofectamine 2000 to produce corresponding viral particles. For cell transfection, chondrocytes were divided into 4 different groups, including the miR-NC group, miR-103a-3p group, siNC and siHMGB1 group according the transfected viral particles. They were transfected with scramble miRNA mimics as miRNA (miR-NC), miR-103a-3p mimics, siNC, and siHMGB1, respectively (purchased from Genepharma, Shanghai, China) using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instruction. Forty-eight hours after incubation, the transfection efficiency was observed using a fluorescence microscope (Olympus Co., Ltd., Beijing, China) and miR-103a-3p expression level was evaluated by quantitative real-time PCR.
Sihmgb1
Manufactured by GenePharma
Sourced in China
The SiHMGB1 is a laboratory equipment product designed for research purposes. It functions as a tool for the analysis and manipulation of the HMGB1 gene, which is involved in various cellular processes. The SiHMGB1 provides researchers with the capability to study the expression and regulation of the HMGB1 gene, enabling them to further their understanding of its role in biological systems.
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Lab products found in correlation
Si nc, by GenePharma (3 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Hbzy 1, by iCell Bioscience (1 mentions)
Oe hmgb1, by GenePharma (1 mentions)
Lipofectamine reagent, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Sirna, by GenePharma (1 mentions)
Mir nc, by GenePharma (1 mentions)
Lipofectamine 2000, by Genechem (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Si hmgb1, by Thermo Fisher Scientific (1 mentions)
Si con, by Thermo Fisher Scientific (1 mentions)
Si con, by GenePharma (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Si pparγ, by GenePharma (1 mentions)
6 protocols using sihmgb1
1
Overexpression of miR-103a-3p in Chondrocytes
2
Mesangial Cell-Based AKI Model
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Rat glomerular mesangial cells (HBZY-1) were purchased from icell bioscience Inc. (iCell-r013) and cultured in deme high glucose medium containing 10% fetal bovine serum, 100 U/ml penicillin and 100 U/ml streptomycin in 5% CO2 incubator at room temperature. Then the cells were seeded onto 24-well plates (5 × 105 cells/well) and divided into seven groups: (1) Control, (2) LPS, (3) LPS + mimics NC, (4) LPS + mimics miR-22, (5) LPS + si-NC, (6) LPS + si-HMGB1, (7) LPS + oe-HMGB1. After the concentration of the cells reached 80%, all the HBZY-1 cells were transfected with mimics miR-22, mimics NC, si-HMGB1, oe-HMGB1, si-NC (GenePharma) for 6 h according to the grouping. Next, except for the control group, all cells were exposed to LPS (10 μg/m) for 12 h to induce AKI. After the transfection, the cell suspension was centrifuged at 800 g/min for 10 min, collected the supernatant and then filtered it with 0.22 μM microporous membrane filtration, collected the filtration solution, and stored it at – 40 ℃ for the following experiments.
3
HMGB1 Plasmid and siRNA Transfection in NP Cells
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The HMGB1 plasmid was constructed by Gene Create (Wuhan, China). An siRNA against HMGB1 (siHMGB1) with a target sequence of 5′-GAGUCCUGGAUGAUACUAATT-3′ and a simulated sequence of 5′-UUCUCCGAACGUGUCACGUTT-3′ was constructed by GenePharma (Shanghai, China). NP cells were transfected using Lipofectamine 3000 (Invitrogen, Carlsbad, USA). Briefly, NP cells were seeded in 6-well plates. After reaching 60%–70% confluence, plasmid (2 μg) or siRNA (150 pmole) was added along with Lipofectamine reagent and Opti-MEM (Gibco). Twelve hours after transfection, the medium was replaced by fresh medium, and regular culture was performed for certain experiments.
4
MiR-548b Regulation of HMGB1 Expression
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MiR-548b mimics, an miR-548b inhibitor, negative control mimics (NC mimics), and a negative control inhibitor (NC inhibitor) were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China), whereas synthetic small interfering RNA (siRNA) against HMGB1 expression (si-HMGB1) and negative control siRNA (si-NC) from Shanghai GenePharma Co., Ltd. (Shanghai, China). HMGB1 plasmid pCMV-HMGB1 and empty plasmid pCMV were synthesized by OriGene (Rockville, MD, USA). Cells were seeded in six-well plates at a density of 4 × 105 cells per well and grown to 70–80% confluence. A cell transfection assay was performed with Lipofectamine™ 2000 (Invitrogen, Thermo Fisher Scientific, Inc.). All the procedures of the transfection assay were based on the product specifications. After transfection for 6 hrs, the cells were washed with phosphate-buffered saline (PBS; Gibco, Thermo Fisher Scientific, Inc.) and cultured in fresh DMEM containing 10% of FBS.
5
Silencing HMGB1 in Breast Cancer
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Small interfering RNA was designed to silence HMGB1 expression. Si‐Con and si‐HMGB1 were synthesized by GenePharma (Shanghai, China). Si‐Con (empty plasmid) and si‐HMGB1 (5'‐GCTCAGACATTGTAGGATT‐3') were transfected into BC cells by Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) for 48 hours at 37°C in accordance with the manufacturer's instructions.
6
Silencing HMGB1 and PPARγ in Cardiomyocytes
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Cardiomyocytes were cultured in serum-free maintenance medium at 37°C with 5% CO2. At 80% confluence, cells were transfected with 100 nM non-targeting small interfering RNA (si)-negative control (NC; siNC), siHMGB1 or siPPARγ (all purchased from Shanghai GenePharma Co., Ltd.) for 48 h at 37°C using Lipofectamine RNAiMAX (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. Following 48 h of transfection, the cells were used for subsequent experiments. The sequences of each siRNA were as follows: siNC, 5′-GCTCTGGAGCAGTTCCGATAT-3′; siHMGB1, 5′-CCATCACAGTGTTGTTAA-3′; and siPPARγ, 5′-TAACGAATGGGATTTGTCTG-3′.
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