pcDNA-FGD5-AS1 overexpression vector and the NC vector were constructed by GenePharma (Shanghai, China). FGD5-AS1/BST2 shRNA (shFGD5-AS1/BST2) or shRNA control (sh-NC), miR-129-5p mimics, inhibitors, or NCs were purchased from GenePharma (Shanghai, China). The Lipofectamine 3000 transfection reagent (Invitrogen, USA) was carried out based on the manufacturer's instruction.
Mir 129 5p mimic
Manufactured by GenePharma
Sourced in China
MiR-129-5p mimic is a laboratory product developed by GenePharma. It is a synthetic oligonucleotide designed to mimic the function of the naturally occurring microRNA miR-129-5p. The core function of this product is to enable the study and investigation of miR-129-5p and its biological roles.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (7 mentions)
Lipopolysaccharides (lps), by Merck Group (2 mentions)
Mimic nc, by GenePharma (2 mentions)
Nc vector, by GenePharma (1 mentions)
Control shrna sh nc, by GenePharma (1 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Mir 410 3p, by GenePharma (1 mentions)
Mir 505 3p, by GenePharma (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ficoll paque, by Cytiva (1 mentions)
Mir 129 5p inhibitor, by GenePharma (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Mir nc, by GenePharma (1 mentions)
Sirna neat1, by GenePharma (1 mentions)
Oe neat1, by GenePharma (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Nc sirna, by GenePharma (1 mentions)
Si malat1, by GenePharma (1 mentions)
Plasmid extraction kit, by Promega (1 mentions)
12 protocols using mir 129 5p mimic
1
Overexpression of FGD5-AS1 in Cells
2
Chondrocyte Overexpression and Inflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For overexpression, miR-410-3p, miR-505-3p, and miR-129-5p mimics (GenePharma) and negative control vectors were added to the culture medium for transfection of cultured chondrocytes using Lipofectamine™ 2000 (Invitrogen). 24 h later, cells were treated with 100 ng/ml lipopolysaccharide (LPS) (Sigma) for 6 h to induce inflammation.
3
Silencing SNHG12 by siRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Small interfering RNAs targeting SNHG12 (si-SNHG12), si-NC (negative control), miR-129-5p mimics, and inhibitors were purchased from GenePharma (Shanghai, China). All of them were transfected using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA).
4
Validation of YWHAB 3'UTR Targeting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
miR-129-5p mimics and miR-129-5p inhibitors were synthesized by Genepharma group (Shanghai, China). The full-length 3'UTR of YWHAB was subcloned into the pIS0 luciferase plasmid to generate pIS0-YWHAB-3'UTR 26 (link). Mutant construct of YWHAB-3'UTR, named pIS0-YWHAB-3'UTR-m, which carried a substitution of three nucleotides within the core binding sites of YWHAB-3'UTR, was conducted using mutant PCR primers. Primers used in this study are shown in Supplementary Table 1 . Lipofectamine 2000 (Life Technologies Corporation, Grand Island, NY, USA) was used for transfection of DNA plasmids and oligonucleotides according to the manufacturer's protocol.
5
Monocyte Isolation and Transfection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As the previous study described, the monocytes were isolated using density gradient centrifugation with FicollPaque (Amersham Pharmacia, Biotech AB) [19 (link)]. The specific cell markers CD14 and CD45 were used to evaluate the purity of the cells, and the purity of the cells was confirmed to be more than 95% by flow cytometry. The extracted monocytes were incubated in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) at 37°C. To mimic the inflammatory condition of NS in vitro, the monocytes were stimulated by 100 ng/ml LPS (Sigma-Aldrich; Merck KGaA) for 4 h.
Monocytes were plated into 48-well plates and lipofectamine 2000 (Thermo Fisher Scientific, Inc.) was used for the transfection according to the manufacturer’s protocols. miR-129-5p mimic (5’-CUUUUUGCGGUCUGGGCUUGC-3’), miR-129-5p inhibitor (5’-GCAAGCCCAGACCGCAAAAAG-3’), and the negative control (miR-NC, 5’-CAGUACUUUUGUGU AGUACAAA-3’) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).
Monocytes were plated into 48-well plates and lipofectamine 2000 (Thermo Fisher Scientific, Inc.) was used for the transfection according to the manufacturer’s protocols. miR-129-5p mimic (5’-CUUUUUGCGGUCUGGGCUUGC-3’), miR-129-5p inhibitor (5’-GCAAGCCCAGACCGCAAAAAG-3’), and the negative control (miR-NC, 5’-CAGUACUUUUGUGU AGUACAAA-3’) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).
6
Ethanol-Induced AML-12 Liver Cell Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse liver cell line AML-12 was purchased from Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China) and cultured in DMEM-F12 medium containing 10% fetal bovine serum (FBS). Upon cell adherence, they were passaged the second day, and those in the logarithmic growth phase were used in the experiments.
AML-12 cells were stimulated by 100 mM ethanol for 24 h alone, or stimulated by 100 mM ethanol for 24 h, followed by transfection of siRNA-NEAT1 negative control (NC), siRNA-NEAT1, miR-129-5p mimic NC, miR-129-5p mimic, NEAT1 overexpression vector (overexpressed (oe)-NEAT1) together with miR-129-5p mimic NC, or oe-NEAT1 together with miR-129-5p mimic. The siRNA-NC, siRNA-NEAT1, mimic-NC, miR-129-5p mimic and oe-NEAT1 were all acquired from GenePharma Co., Ltd, (Shanghai, China).
AML12 cells were seeded in 12-well plates and cultured for 24 h before transfection. The transfection was conducted by Lipofectamine 2000 reagent (Invitrogen Inc., Carlsbad, CA, USA) when the cell confluence reached 70–90%. Transfected for 6 h, the medium was changed and cells were stimulated by 100 mM ethanol for 24 h. Finally, cells and cell supernatant were collected for the subsequent cell experiments.
AML-12 cells were stimulated by 100 mM ethanol for 24 h alone, or stimulated by 100 mM ethanol for 24 h, followed by transfection of siRNA-NEAT1 negative control (NC), siRNA-NEAT1, miR-129-5p mimic NC, miR-129-5p mimic, NEAT1 overexpression vector (overexpressed (oe)-NEAT1) together with miR-129-5p mimic NC, or oe-NEAT1 together with miR-129-5p mimic. The siRNA-NC, siRNA-NEAT1, mimic-NC, miR-129-5p mimic and oe-NEAT1 were all acquired from GenePharma Co., Ltd, (Shanghai, China).
AML12 cells were seeded in 12-well plates and cultured for 24 h before transfection. The transfection was conducted by Lipofectamine 2000 reagent (Invitrogen Inc., Carlsbad, CA, USA) when the cell confluence reached 70–90%. Transfected for 6 h, the medium was changed and cells were stimulated by 100 mM ethanol for 24 h. Finally, cells and cell supernatant were collected for the subsequent cell experiments.
7
Modulating MALAT1 and miR-129-5p in GSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
si-MALAT1, miR-129-5p mimic, inhibitor, and normal control (NC) were manufactured by Genepharma Company (Shanghai, China) and transfected into GSCs and microglia cells at exponential phase (40–50% confluence, 50 nm) with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) on a six-well plate, which were further incubated for 48 h to extract RNA/protein. All of these performances were followed by the protocol recommended by the manufacturer.
8
Investigating miR-129 Function in H9C2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
miR-129-5p mimic and negative controls (NC) miRNA were synthesized from Shanghai Gene Pharma Co., Ltd. (China). To investigate the function of miR-129 in a cardiomyocyte, H9C2 cells were transfected with miR-129 mimics (final concentration100 nm) by Lipofectamine 2000 (Ruibo, China).
9
Regulation of SOX6 by miR-129-5p
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The targeting relationship between miR-129-5p and SOX6 and the binding site between miR-129-5p and SOX6 3ʹuntranslated region (UTR) were forecasted by bioinformatics software http://www.targetscan.org . The 3ʹUTR fragment of SOX6 gene was amplified by PCR and cloned into pmirGLO vector to construct the recombinant luciferase reporter plasmid of wild type plasmid (SOX6-WT) and mutant type plasmid (SOX6-MUT). SOX6-WT and SOX6-MUT plasmids were extracted according to the steps of the purchased plasmid extraction kit (Promega, Madison, Wisconsin, USA). The logarithmic cells were inoculated into 96-well plates. When the cell confluence was about 70%, Lipofectamine 2000 was utilized for transfection. SOX6-WT and SOX6-MUT were mixed with mimic NC and miR-129-5p mimic (Shanghai GenePharma Co. Ltd., Shanghai, China) respectively, and then co-transfected to neuronal cells. The cells were amassed and lysed after transfected 48 h, and luciferase activity was detected by luciferase detection kit (BioVision, San Francisco, CA, USA) and Glomax 20/20 luminometer (Promega, Madison, Wisconsin, USA).
10
Modulating miR-129-5p and SOX6 in Alzheimer's Disease
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were divided into 8 groups: control group (normal rat hippocampal neuron cell); AD group (AD rat hippocampal neuron cell); mimic NC group (AD rat hippocampal neuron cells transfected with mimic-NC); miR-129-5p mimic group (AD rat hippocampal neuron cells transfected with miR-129-5p mimic, GeneCopoeia Co., Ltd. Guangzhou, China); siRNA-NC group (AD rat hippocampal neuron cells transfected with SOX6-siRNA vector NC); SOX6-siRNA group (AD rat hippocampal neuron cells transfected with SOX6-siRNA vector, GenePharma Ltd. Company, Shanghai, China); miR-129-5p mimic + OE-NC group (AD rat hippocampal neuron cells transfected with miR-129-5p mimic and overexpression of SOX6 vector NC); miR-129-5p mimic + OE-SOX6 group (AD rat hippocampal neuron cells transfected with miR-129-5p mimic and overexpression SOX6 vector, GeneCopoeia Co., Ltd. Guangzhou, China). The cells were inoculated in six-well plate before 24 h with transfection. When the confluence reached about 50%, neuronal cells were transiently transfected under the mediation of lipofectamine 2000 (Invitrogen, Carlsbad, California, USA), transfected for 6 h, and collected for subsequent experiments after cultured for 48 h.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!