To determine the potential function of miR-144-3p, RAW264.7 cells were transiently transfected with 50 nM control or miR-144-3p mimic (GenePharma, Shanghai, China); 50 nM control or miR-144-3p inhibitor (GenePharma, Shanghai, China) by using Lipofectamine 2000 (Invitrogen, USA).
Mir 144 3p inhibitor
Manufactured by GenePharma
Sourced in China
The MiR-144-3p inhibitor is a laboratory reagent designed to inhibit the expression of the miR-144-3p microRNA. It is commonly used in research applications to study the functional role of miR-144-3p in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Mir 144 3p mimic, by GenePharma (4 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Inhibitor nc, by GenePharma (3 mentions)
Mimic nc, by GenePharma (2 mentions)
Mcf 10a, by Cobioer Biosciences (1 mentions)
Mda mb 157, by Cobioer Biosciences (1 mentions)
Si linc00461, by GenePharma (1 mentions)
Lipofectamin 2000, by Thermo Fisher Scientific (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Si nc, by GenePharma (1 mentions)
Oe nc, by GenePharma (1 mentions)
Mda mb 231, by Cobioer Biosciences (1 mentions)
Mcf 7, by Cobioer Biosciences (1 mentions)
H460 ddp, by Thermo Fisher Scientific (1 mentions)
Pc xist, by GenePharma (1 mentions)
A549 ddp, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
5 protocols using mir 144 3p inhibitor
1
Functional Evaluation of miR-144-3p
2
Breast Cell Line Transfection Protocol
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Human normal breast epithelial cell line MCF 10A (CBP60419) and breast cancer cell lines AU565 (CBP60353), MCF-7 (CBP60380), MDA-MB-157 (CBP60381) and MDA-MB-231 (CBP60382) were all purchased from Cobioer (Nanjing, China). Plasmids including si-NC, si-LINC00461, inhibitor NC, miR-144-3p inhibitor, mimic NC, miR-144-3p mimic, oe-NC, oe-KPNA2 were ordered from GenePharma (Shanghai, China), and the siRNAs were sequenced as below: si-LINC00461: 5′-CTGCAAAGAAGCATAAAATGA-3′; si-NC: 5′-TTCTCCGAACGTG TCACGT-3′.
Before transfection, cells were grown in 6-well plates (3 × 105 cells/well) until the density reached 50%. 250 μL of serum-free Opti-MEM (51985042, Gibco, Gaitherburg, MD, USA) was used to dilute the target plasmids (4 μg) and Lipofectamin 2000 reagent (10 μL; 11668-019, Invitrogen, NY, California, USA), respectively. Thereafter, the dilutions were mixed, and then dripped into cells after 20 min. All cells were maintained in 5% CO2 at 37 °C for 6 h, and cultured for additional 36–48 h with fresh mediums.
Before transfection, cells were grown in 6-well plates (3 × 105 cells/well) until the density reached 50%. 250 μL of serum-free Opti-MEM (51985042, Gibco, Gaitherburg, MD, USA) was used to dilute the target plasmids (4 μg) and Lipofectamin 2000 reagent (10 μL; 11668-019, Invitrogen, NY, California, USA), respectively. Thereafter, the dilutions were mixed, and then dripped into cells after 20 min. All cells were maintained in 5% CO2 at 37 °C for 6 h, and cultured for additional 36–48 h with fresh mediums.
3
Insulin-Producing Cell Line Manipulation
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832/3 rat insulinoma cell line (INS-1) purchased from Sigma-Aldrich (MO, USA) were grown in DMEM medium containing 10% FBS and antibiotics at 37 °C in a CO2 incubator.
Mimic and inhibitor for miR-144-3p (miR-144-3p mimic and miR-144-3p inhibitor) and the corresponding negative controls (NC mimic and NC inhibitor) were supplied by the GenePharma company (Shanghai, China). Gene-specific (USP22 or SIRT1) overexpression plasmids or shRNAs suppling by the GenePharma were transfected into cells to overexpress or knock down the expression level of proteins. According to standard protocols, INS-1 cells were transfected with the stated plasmids using Lipofectamine 2000 (Life Technologies, CA, USA) and harvested at 48 h post transfection for further experiments.
Mimic and inhibitor for miR-144-3p (miR-144-3p mimic and miR-144-3p inhibitor) and the corresponding negative controls (NC mimic and NC inhibitor) were supplied by the GenePharma company (Shanghai, China). Gene-specific (USP22 or SIRT1) overexpression plasmids or shRNAs suppling by the GenePharma were transfected into cells to overexpress or knock down the expression level of proteins. According to standard protocols, INS-1 cells were transfected with the stated plasmids using Lipofectamine 2000 (Life Technologies, CA, USA) and harvested at 48 h post transfection for further experiments.
4
Investigating Cisplatin Resistance in NSCLC
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Cisplatin-resistant human NSCLC cell lines, H460/DDP and A549/DDP, were obtained from RiboBio Co. (Guangzhou, China). Cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco, Grand Island, NY, USA) with 10% fetal bovine serum (FBS) (HyClone, Logan, UT, USA), penicillin and and streptomycin (Sigma-Aldrich, St. Louis, MO, USA) at 37°C with 5% CO2. Culture medium was changed every 2 days.
The negative control (NC) mimic, microRNA (miR)-144-3p mimic, NC inhibitor and miR-144-3p inhibitor were purchased from GenePharma Co., Ltd. (Shanghai, China). Short-hairpin (sh)-NC, sh-XIST, pc-NC and pc-XIST were constructed by GenePharma Co., Ltd. (Shanghai, China). All vectors and oligonucleotides were transfected into H460/DDP and A549/DDP cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s instructions.
The negative control (NC) mimic, microRNA (miR)-144-3p mimic, NC inhibitor and miR-144-3p inhibitor were purchased from GenePharma Co., Ltd. (Shanghai, China). Short-hairpin (sh)-NC, sh-XIST, pc-NC and pc-XIST were constructed by GenePharma Co., Ltd. (Shanghai, China). All vectors and oligonucleotides were transfected into H460/DDP and A549/DDP cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s instructions.
5
miR-144-3p Regulation of ERO1L Expression
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The miR-144-3p mimics (5'-UACAGUAUAGAUGAUGUACU-3'), mimics negative control (5'-UUCUCCGAACGUGUCACGUTT-3'), miR-144-3p inhibitor (5'-AGUACAUCAUCUAUACUGUA-3'), and inhibitor negative control (5'-CAGUACUUUUGUGUAGUACAA-3') were purchased from GenePharma (Shanghai, China). The ERO1L overexpression plasmid was constructed by introducing the human ERO1L cDNA into the pcDNA3.1 vector (Invitrogen), and was designated as pcDNA3.1-ERO1L. Specific siRNA against ERO1L (5'-GCACTGCTCTGAAGATCTT-3') and a scramble siRNA (5'-TTCTCCGAACGTGTCACGT-3') were synthesized by RiboBio (Guangzhou, China). Cells were seeded in 12-well plates and transfected with the above oligonucleotides or vectors using Lipofectamine 3000 (Invitrogen) following the manufacturer's instructions. Cells were harvested at 72 h after transfection and used in subsequent analyses.
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