The HT‐29 and Caco‐2 colonic cell lines were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). MiR‐124‐3p mimics, miR‐96‐5p mimics, miR‐124‐3p inhibitors and miR‐96‐5p inhibitors were purchased from GenePharma (Shanghai, China), and the sequences are shown in Table S1 . Knockdown or overexpression lentivirus vectors (Lv‐shRab27A, Lv‐shSTAT3, Lv‐Rab27A and Lv‐STAT3) were purchased from GenePharma (Shanghai, China). The cells were distributed in 6‐well plates to approximately 50%‐70% confluence and were transfected the next day with plasmid at a concentration of 100 nmol\L in DMEM (GenePharma, Shanghai, China) using Lipofectamine 3000 (Invitrogen, USA), according to the manufacturer's instructions.
Mir 124 3p mimic
Manufactured by GenePharma
Sourced in China, United States
MiR-124-3p mimic is a synthetic RNA molecule designed to mimic the function of the naturally occurring microRNA, miR-124-3p. MicroRNAs are small, non-coding RNA molecules that play a crucial role in the regulation of gene expression. The MiR-124-3p mimic can be used as a research tool to study the biological functions and mechanisms associated with miR-124-3p in various cellular and molecular processes.
Automatically generated - may contain errors
Lab products found in correlation
Mir 124 3p inhibitor, by GenePharma (7 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (6 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Sh nc, by GenePharma (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Mir 96 5p mimic, by GenePharma (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000, by GenePharma (1 mentions)
Si hoxa11 as, by GenePharma (1 mentions)
Empty pcdna3, by GenePharma (1 mentions)
Mimic nc, by GenePharma (1 mentions)
Mir nc, by GenePharma (1 mentions)
Pcdna xist, by GenePharma (1 mentions)
Sh xist, by GenePharma (1 mentions)
Sh irf1, by GenePharma (1 mentions)
Pmirglo dual luciferase reporter gene vector, by Promega (1 mentions)
Si neat1, by GenePharma (1 mentions)
Neat1 overexpression vector, by GenePharma (1 mentions)
Pcdna empty vector pcdna, by GenePharma (1 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
14 protocols using mir 124 3p mimic
1
Regulation of Colonic Cell Lines by miRNA and Protein Knockdown
2
Tracking Exosomal miR-124-3p Uptake in Microglia
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Neurons were transfected with 5′-carboxyfluorescein (FAM)-labeled miR-124-3p mimics, miR-124-3p inhibitor, and their corresponding negative controls (GenePharma) with Lipofectamine 3000. After that, exosomes were extracted from the culture medium, divided into four different groups, and added into target microglial cells. After co-incubation, microglial cells were fixed with 4% PFA, permeabilized with 0.05% Trition X-100, and stained with DAPI (Thermo Fisher Scientific). Images were acquired using a confocal microscope to observe green signaling intensity in the target microglial cells.
3
HOXA11-AS Regulation of miR-124-3p
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Cells in logarithmic growth phase were taken, and then seeded in 6-well plates at 5×106/well after trypsinization. The cells were transfected after the cell growth was stable. HOXA11-AS over-expression plasmid (HOXA11-AS), si-HOXA11-AS, miR-124-3p mimics and their negative controls were obtained from GenePharma (Shanghai, China). The cells were incubated at 37° C with 5% CO2, and were transfected with Lipofectamine® 3000 (Invitrogen; ThermoFisher Scientific, Inc.) reagent according to the instructions of FuGENE® HD Transfection Reagent (Roche, Shanghai, China). After 24 hours of transfection, total cellular RNA was extracted for real-time fluorescence quantitative PCR to monitor the changes in miR-124-3p and HOXA11-AS expressions in the transfected cells.
4
Transfection of HRMECs with Plasmids
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The pcDNA3.1/G3BP2 and its negative control (empty pcDNA3.1), miR-124-3p mimics and its negative control (NC mimics) were purchased from GenePharma (Shanghai, China). The above-mentioned vectors were transfected into HRMECs for 24 h using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, United States) according to the manufacturer’s instructions.
5
Hypoxia/Reoxygenation-Induced NRK-52E Cell Transfection
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The pcDNA3.1 XIST (pcDNA-XIST), pcDNA-NC, miR-124-3p mimics, miR-NC, short hairpin- (sh-) ITGB1, and sh-NC were synthesized by GenePharma (Shanghai, China). H/R-treated NRK-52E cells grown to 85% confluence were transfected or cotransfected with these above agents using Lipofectamine 3000 (Invitrogen). The H/R-treated NRK-52E cells in the blank group did not receive any transfection.
6
PDIA3P1 Plasmid Overexpression and Regulation
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Human full length PDIA3P1 plasmids was selected and inserted into the pLVX-IRES-Puro vector for stable overexpression. The si-PDIA3P1, si-RELA, miR-124-3p mimics, miR-124-3p inhibitors, and negative controls were purchased from Genepharma (Shanghai, China). All sequences are listed in Supplementary Table 2 . Plasmids were purchased from Bioscience (Jinan, China), and the human PDIA3P1 promoter sequence was obtained from the UCSC Genome Browser (http://genome.ucsc.edu/ ). For transfection, cells were seeded in 6-well plates overnight and transfected using Lipofectamine 3000 (Invitrogen, CA, USA) according to the manufacturer’s instructions. Cells were harvested for RT-qPCR and western-blot analysis 48 h after transfection.
7
Transfection of BV2 Cells with XIST, miR-124-3p, and IRF1 Modulators
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PcDNA-XIST (OV-XIST group), sh-XIST (KD-XIST group), miR-124–3p mimic, miR-124–3p inhibitor, sh-IRF1 and the corresponding negative control were synthesized by Genepharma (Shanghai, China). The above-mentioned plasmids were separately transfected into BV2 cells through Lipofectamine 3000 (Invitrogen, CA, USA). After 48 h, real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) or Western blot assay was utilized to assess the efficiency of transfection.
8
Validation of miR-124-3p and ANGPTL2 Interaction
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The binding sites of miR-124-3p and ANGPTL2 were predicted using the StarBase database (https://starbase.sysu.edu.cn/ ). Subsequently, the complementary binding sequences and mutated sequences of miR-124-3p and ANGPTL2 were amplified and cloned into the pmiRGLO dual-luciferase reporter gene vector (Promega, Madison, WI, USA) to construct wild-type plasmid pmiRGLO-ANGPTL2-WT and the corresponding mutant plasmid pmiRGLO-ANGPTL2-MUT. In accordance with the instructions of LipofectamineTM 2000 (Invitrogen), the plasmid was cotransfected with mimic NC or miR-124-3p mimic (sequence: 5′-UAAGGCACGCGGUGAAUGCCCA-3′, purchased from GenePharma, Shanghai, China) into ovarian granulosa cells KGN (Sunncell, Wuhan, Hubei, China). Afterwards, the luciferase activity was detected 48 h later.
9
Modulating NEAT1 and PDE4B in Parkinson's Disease
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Small interfering RNA (siRNA) against NEAT1 (si-NEAT1), siRNA against PDE4B (si-PDE4B), siRNA negative control (si-NC), NEAT1 overexpression vector, PDE4B overexpression vector, pcDNA empty vector (pcDNA), miR-124-3p mimic, mimic negative control (miR-NC), miR-124-3p inhibitor, and inhibitor control (NC inhibitor) (sequences are detailed in Table 1 ) were available from GenePharma (Shanghai, China). Cell transfection was carried out with Lipofectamine™ 2000 transfection reagent (Invitrogen) 48 h before MPP+ treatment.
The sequences of the miRNAs used in this study
| Targets | Sequence |
|---|---|
| 5ʹ-UAAGGCACGCGGUGAAUGCCCA-3’ | |
| 5ʹ-UUCUCCGAACGUGUCACGUTT-3ʹ | |
| 5ʹ-GGCAUUCACCGCGUGCCUUA-3’ | |
| 5ʹ-CAGUACUUUUGUGUAGUACA-3ʹ |
10
Investigating miR-124-3p Targeting of IRE1α
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A luciferase reporter analysis was conducted to investigate whether miR-124-3p directly targets IRE1α mRNA in order to detect miR-124-3p targets related to ER stress. IRE1α luciferase reporter plasmids were constructed by inserting the IRE1α 3′-untranslated region (3′UTR), including all predicted miR-124-3p binding sites, into the pGL3 luciferase reporter vector. In short, the IRE1α fragment was PCR-amplified from mouse genomic DNA and ligated into the pGL3 control vector downstream of the SV40 promoter using the XbaI sites (Promega, Madison, WI, USA). To construct a mutant (MUT) version of the plasmid, the miR-124-3p binding site was removed.
As we reported previously (Li et al., 2019), HT22 neuronal cells were seeded in 96-well plates. Employing Lipofectamine 2000, the miR-124-3p mimic or random oligonucleotides (Gene Pharma) were cotransfected into neurons with the wild-type or mutant pGL3-IRE1α-3′UTR. Following incubation for 2 days, the cells were harvested, and luciferase activity was detected using a Dual-Luciferase Reporter Assay System (Promega).
As we reported previously (Li et al., 2019), HT22 neuronal cells were seeded in 96-well plates. Employing Lipofectamine 2000, the miR-124-3p mimic or random oligonucleotides (Gene Pharma) were cotransfected into neurons with the wild-type or mutant pGL3-IRE1α-3′UTR. Following incubation for 2 days, the cells were harvested, and luciferase activity was detected using a Dual-Luciferase Reporter Assay System (Promega).
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