Small interfering RNA (siRNA) targeting APJ (siRNA-APJ, sequence: CTGACATGTTACTTCTTCA), siRNA-APJ negative control (NC), miR-124-3p mimic, NC mimic, miR-124-3p inhibitor, and NC inhibitor were synthesized by RiboBio Co. (Guangzhou, China). CTDSP1 overexpression adenovirus and the CTDSP1 overexpression adenovirus negative control (NC) were synthesized by GeneChem. (Shanghai, China). Neurons were transfected with siRNA-APJ, siRNA-APJ NC, miR-124-3p mimic, NC mimic, miR-124-3p inhibitor, NC inhibitor, CTDSP1 overexpression adenovirus, or CTDSP1 overexpression adenovirus NC using transfection reagent according to the manufacturer’s instructions (RiboBio Co, China). During this period, neurons were incubated in culture medium without the penicillin/streptomycin solution.
Mir 124 3p mimic
Manufactured by RiboBio
Sourced in China
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 role in the regulation of gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Mir 124 3p inhibitor, by RiboBio (3 mentions)
Mimic nc, by RiboBio (2 mentions)
Sicontrol, by Thermo Fisher Scientific (2 mentions)
Inhibitor nc, by RiboBio (1 mentions)
Sirna apj, by RiboBio (1 mentions)
Si control, by RiboBio (1 mentions)
Lipofectamine 2000, by RiboBio (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Eca 109, by Thermo Fisher Scientific (1 mentions)
Kyse 150, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
5 azacitidine, by Merck Group (1 mentions)
Dnmt1 sirna, by RiboBio (1 mentions)
Kyse 150, by Merck Group (1 mentions)
Eca109, by Merck Group (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Mir nc, by RiboBio (1 mentions)
Si snhg14, by RiboBio (1 mentions)
9 protocols using mir 124 3p mimic
1
Modulating Neuronal APJ and miR-124-3p
2
Transfection of miR-124-3p Mimics and Inhibitors
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Small interfering RNA (siRNA), si-control, miR-124-3p mimic, miR-124-3p miR-124-3p inhibitor, negative control were purchased from Ribobio (Guangzhou, China). For transfection, miR-124-3p mimic, miR-124-3p inhibitor, negative control, siRNA or si-control in Lipofectamine 2000 (Invitrogen) was transfected into cells according to the manufacturer’s instructions. The siRNA sequences are listed in Additional file 2 : Table S3.
3
miR-124-3p Regulation of lncRNA RP11-395G23.3
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miR-124-3p mimic, miRNA negative control (miR-NC), and miR-124-3p inhibitor were purchased from RiboBio (RiboBio, Guangzhou, China). Small interfering RNAs (siRNAs) against RP11-395G23.3 (si-RP11-395G23.3) and negative control siRNA (siControl) were purchased from Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). The sequences were: siControl: 5′-UUCUCCGAACGUGUCACGUTT-3′; si-RP11-395G23.3: 5′-GGAGUUCUCCACAUGUAAATT-3′. They were transfected into Nthy-ori 3-1, C643, and HTh-7 cells by Lipofectamine 2000 (Invitrogen, Thermo Fisher Scientific, Inc.), according to the manufacturer’s protocols.
4
NLRP3 Inflammasome Regulation in Alzheimer's Disease
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NC mimic, miR-124-3p mimic, NC inhibitor and miR-124-3p inhibitor, lentivirus vector (LV) – MALAT1 and shRNA-MALAT1 were purchased from Ribobio Co., Ltd. (Guangzhou, China). To investigate the role of NLRP3 inflammasome activation in AD, CD4+T cells were co-treated with NLRP3 activator MSU (0.5 mg/mL) and icariin (100 µM) or treated with NLRP3 inhibitor MCC950 (10 µM) alone, and all cells were cultured in Th2-condition. To investigate the molecular regulatory mechanism of icariin in Th2 skewed cells, CD4+T cells were transfected with the indicated transfectants for 24 h to obtain stably transfected cells, then treated with or without 100 µM icariin and cultured in Th2-cell condition.
5
Extrinsic miR-124-3p Transfection in Colon Cancer Cells
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According to the manufacturer's instructions, 10 × 104 LS174 T and HT‐29 cells were seeded on the 6‐well plates. Extrinsic miR‐124‐3p mimic (Cat.No.miR10000422‐1‐5, RiboBio, China) or the respective NC (Cat.No.miR01101‐1‐5) was transiently transfected with Lipofectamine 2000 (Cat.No.11668019, Invitrogen, USA) for 6 h and replaced with culture medium for another 72 or 96 h. Transfected cells were used for further analysis.
6
miR-124-3p Transfection in ADSCs
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The miR-124-3p mimics and its corresponding negative control (NC) were obtained from RiboBio (Guangzhou, China) and were transfected into adipose stem cells (ADSCs) cells utilizing Lipofectamine 2000 reagents (Invitrogen, Carlsbad, CA, USA), following the supplier’s instructions. The concentration of transfection was 50 nm, and the solution was altered after 6 hours transient transfer. After 48 hours, the cells were subjected to qPCR for verification.
7
Esophageal Cell Line Maintenance and Manipulation
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Human normal esophageal cell line HEEC (Het-1A, ATCC® CRL-2692™) was obtained from the America Type Culture Collection in May 2017, and two human esophageal carcinoma cell lines, KYSE-150 (TCHu236) and Eca-109 (TCHu69), were obtained from Shanghai Institute of Biological Sciences, Chinese Academy of Sciences in May 2017. These cell lines have been authenticated by short-tandem repeat analyses. They are free of mycoplasma contamination.
The cells were maintained in RPMI-1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% FBS (Gibco, Grand Island, NY, USA) and 1% Penicillin-Streptomycin (10,000 U/mL) (Invitrogen). For cell transfection, siRNAs including DNMT1 siRNA, BCAT1 siRNA, scrambled siRNA (NC siRNA), miR-124-3p mimics and its negative control (NC mimics) were supplied by Ribobio Company (Guangzhou, China) and pcDNA-BCAT1(OE-BCAT1) or pcDNA-negative (OE-NC) were constructed using pcDNA3.1(+) vector. KYSE-150 and Eca-109 cells grown in 6-well cell plates (1 × 106) were transfected with siRNA or pcDNA3.1vector using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instruction. After 6 h, the transfection solution was replaced by complete medium.
For drug treatment, KYSE-150 or Eca109 cells were treated with 5 μM 5-Azacitidine (5-Aza, Sigma-Aldrich, Saint Louis, MO, USA), which is the inhibitor of DNMT1.
The cells were maintained in RPMI-1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% FBS (Gibco, Grand Island, NY, USA) and 1% Penicillin-Streptomycin (10,000 U/mL) (Invitrogen). For cell transfection, siRNAs including DNMT1 siRNA, BCAT1 siRNA, scrambled siRNA (NC siRNA), miR-124-3p mimics and its negative control (NC mimics) were supplied by Ribobio Company (Guangzhou, China) and pcDNA-BCAT1(OE-BCAT1) or pcDNA-negative (OE-NC) were constructed using pcDNA3.1(+) vector. KYSE-150 and Eca-109 cells grown in 6-well cell plates (1 × 106) were transfected with siRNA or pcDNA3.1vector using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instruction. After 6 h, the transfection solution was replaced by complete medium.
For drug treatment, KYSE-150 or Eca109 cells were treated with 5 μM 5-Azacitidine (5-Aza, Sigma-Aldrich, Saint Louis, MO, USA), which is the inhibitor of DNMT1.
8
Regulation of HK-2 Cell Transcripts
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The small interfere- (si-) NC, si-SNHG14, miR-124-3p inhibitor, miR-124-3p mimics, miR-NC, and pcDNA-MMP2 were bought from RiboBio. When HK-2 cells grew 80%-90% confluence, the above transcripts were transfected into HK-2 cells with Lipofectamine 3000 (Invitrogen) for 48 h.
9
Exosomal delivery of miR-124-3p
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EXOs were loaded with miR-124-3p through electroporation using a Gene Pulser Xcell (Bio-Rad Laboratories, Inc.) electroporation system. First, miR-124-3p mimics (Guangzhou RiboBio Co., Ltd.) were mixed with EXOs in an electroporation buffer at a final concentration of 200 nM. The mixtures were then cultured at 4°C for 10 min and transferred to ice-cold 0.2-cm cuvettes for electroporation at 400 V and 125 µF capacitance with a single pulse. Subsequently, the EXOs were maintained on ice for ≥15 min following electroporation. Negative control (NC) mimics (Guangzhou RiboBio Co., Ltd.) mixed with EXOs were electroporated to serve as a control.
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