P3 BMSCs were seeded onto 12‐well plates and transfected based on directions of Lipofectamine 2000 reagent (Invitrogen Inc) when cell confluence reached 80%. MiR‐206 mimic, miR‐206 inhibitor and their NCs were acquired from GenePharma Co., Ltd.. BMSCs were divided into 4 groups: the blank group (no treatment), the NC group (transfection of irrelevant control), the MiR‐206 mimic group (transfection of MiR‐206 mimic) and the miR‐206 inhibitor group (transfection of miR‐206 inhibitor). Cells in each group that had been transfected for 72 hours were used in subsequent experiments.
Mir 206 inhibitor
Manufactured by GenePharma
Sourced in China
The MiR-206 inhibitor is a laboratory instrument designed for the detection and inhibition of the microRNA-206 (miR-206) molecule. It is a tool used in scientific research to study the functions and regulations of this specific microRNA, which has been associated with various biological processes and diseases.
Automatically generated - may contain errors
Lab products found in correlation
Mir 206 mimic, by GenePharma (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Inhibitor nc, by GenePharma (3 mentions)
Mir nc, by GenePharma (2 mentions)
Scrambled mirna, by GenePharma (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Dmem f12 1 1 medium, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Lentivirus, by Genechem (1 mentions)
Mir 206 agomir, by RiboBio (1 mentions)
Pcdna3.0 vector, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Pcdna3.1 malat1, by RiboBio (1 mentions)
Lipofectamine 3000 transfection kit, by Thermo Fisher Scientific (1 mentions)
Mimic nc, by GenePharma (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Hct116, by GenePharma (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Mirna inhibitor, by GenePharma (1 mentions)
11 protocols using mir 206 inhibitor
1
Transfection of miR-206 in BMSCs
2
Modulating miR-206 Expression in Nasopharyngeal Carcinoma Cells
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CNE-1 cells were obtained from Institute of Virology, Chinese Academy of Preventive Medicine; NP-69 (the SV40 large T immortalized nasopharyngeal epithelial cell line) and CNE-2 cells were purchased from the Shanghai Institute of Cell Biology (Shanghai, P.R. China). The EBV+ C666-1 cells were from the cell bank of Xiangya Central Laboratory (Central South University, Changsha, P.R. China). Cells were maintained in RPMI-1640 or DMEM/F12 (1:1) medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing streptomycin, penicillin, and supplemented with 10% fetal bovine serum (Gibco, Thermo Fisher Scientific) and cultured at 37°C in a humidified incubator with 5% CO2. For functional analysis, C666-1 cells grown to 80%–90% confluence were transfected with the miR-206 mimic, the miR-206 inhibitor, or scrambled miRNA as a control (Genepharma Company, Suzhou, P.R. China) using Lipofectamine RNAimax (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). Cells were harvested 48–72 h after transfection for further experiments. Lentiviral transduction of C666-1 cells was carried out with lentivirus carrying miR-206 precursor and its control according to the manufacturer’s protocol (Genechem Company, Shanghai, P.R. China). Stable miR-206-overexpressing cells and the control cells were then selected using puromycin.
3
Modulating miR-206 and PTP1B in Cardiac Cells
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For the miR-206 knockdown, miR-206 overexpression, and PTP1B up-regulation, 100 nM of miR-206 inhibitor (purchased from GenePharma), 50 nM of miR-206 mimics (purchased from GenePharma), and 2 μg of p-EZ-M02-PTP1B plasmid (purchased from FulenGen), respectively, was added to the culture medium for transfection using Lipofectamine™ 2000 (Invitrogen) following the manufacturer’s protocol.
For the overexpression of miR-206 in vivo, miR-206 agomir (RiboBio) was introduced into rat hearts using the procedure described by Hajjar et al. [26 (link)]. In brief, rats were anesthetized and the pericardium was opened. Aorta and pulmonary arteries were identified and followed with aortic clamping. A catheter was advanced to the aortic root through the apex of the left ventricle (LV). And a 200-μl solution that contains 10 nM control or miR-206 agomir (RiboBio) suspension in sterile PBS was injected through the catheter. Aortic clamping remained so the solution was circulated and the heart was perfused. Clamps were released after 10 s and the chest was closed.
For the overexpression of miR-206 in vivo, miR-206 agomir (RiboBio) was introduced into rat hearts using the procedure described by Hajjar et al. [26 (link)]. In brief, rats were anesthetized and the pericardium was opened. Aorta and pulmonary arteries were identified and followed with aortic clamping. A catheter was advanced to the aortic root through the apex of the left ventricle (LV). And a 200-μl solution that contains 10 nM control or miR-206 agomir (RiboBio) suspension in sterile PBS was injected through the catheter. Aortic clamping remained so the solution was circulated and the heart was perfused. Clamps were released after 10 s and the chest was closed.
4
miR-206 Regulation of c-Met Expression
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miR-206 mimics, miR-206 inhibitor, and their negative controls (NCs) were obtained from Shanghai GenePharma Co., Ltd. (Shanghai, China). The coding domain sequences of c-Met mRNA were amplified by PCR, and inserted into a pcDNA 3.0 vector (Invitrogen; Thermo Fisher Scientific, Inc.) to enhance its expression, and was termed pcDNA-c-Met. Eca109 and KYSE410 cells (5.0×105/well) were seeded and cultured to 60–80% confluence in six-well plates. According to the manufacturer's protocol, they were transiently transfected with the 20 nM interference oligonucleotide which included negative control (inhibitor NC: 5′-CAGUACUUUUGUGUAGUACAA-3′; mimics NC: 5′-UUCUCCGAACGUGUCACGUTT-3′), miR-206 inhibitor (5′-CCACACACUUCCUUACAUUCCA-3′) or miR-206 mimics (5′-UGGAAUGUAAGGAAGUGUGUGG-3′) using Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.). Following 6 h transfection at 37°C, the medium was replaced with RPMI1640 media containing 10% FBS and the cells were harvested for further experiments after 48 h.
5
Endometrial Stromal Cell Transfection Protocol
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The primary endometrial stromal cells (ESCs) were obtained according to previous description. ESCs were cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco, USA) with 10% fetal bovine serum (FBS, Gibco, USA), 10 U/ml penicillin, and 100U/ml streptomycin at 37°C and 5% CO2.
ESCs (1х105 cells/mL) were cultured in a 24-well plate for 24 h. The miR-206 mimic, NC-mimic, miR-206 inhibitor, and NC-inhibitor were obtained from GenePharma (Shanghai, China). pcDNA3.1-MALAT1 and pcDNA3.1-MALAT1 negative control (pcDNA-NC) were obtained from RIBO Bio (Guangzhou, China). Lipofectamine 3000 transfection kit (Invitrogen, USA) was used for cell transfection according to the transfection instructions.
ESCs (1х105 cells/mL) were cultured in a 24-well plate for 24 h. The miR-206 mimic, NC-mimic, miR-206 inhibitor, and NC-inhibitor were obtained from GenePharma (Shanghai, China). pcDNA3.1-MALAT1 and pcDNA3.1-MALAT1 negative control (pcDNA-NC) were obtained from RIBO Bio (Guangzhou, China). Lipofectamine 3000 transfection kit (Invitrogen, USA) was used for cell transfection according to the transfection instructions.
6
Establishment of 5-FU Resistant CRC Cell Lines
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CRC cell lines including HCT116 and RKO were obtained from American Type Culture Collection (Rockville, MD, USA). The cells were cultured in Dulbecco’s Modified Eagle’s Medium (Gibco, Gaithersburgh, MD, USA) supplemented with 10% fetal bovine serum (Gibco), and 1% penicillin–streptomycin at 37°C in a humidified environment with 5% CO2. miR-206 inhibitor, miR-206 mimic, or the appropriate negative controls (NCs) of miRNA inhibitor and miRNA mimic were obtained from GenePharma (Shanghai, People’s Republic of China).
5-FU-resistant cell lines, HCT116/FR and RKO/FR, were derived by incubation with stepwise increasing concentrations of 5-FU. Briefly, HCT116 and RKO cells were treated with increasing dose of 5-FU from 10 to 100 μg/mL for more than 12 months. During the treatment, after each round, when the surviving cells reached >70% confluence they were passaged by trypsinization, and the dose of 5-FU was increased.
5-FU-resistant cell lines, HCT116/FR and RKO/FR, were derived by incubation with stepwise increasing concentrations of 5-FU. Briefly, HCT116 and RKO cells were treated with increasing dose of 5-FU from 10 to 100 μg/mL for more than 12 months. During the treatment, after each round, when the surviving cells reached >70% confluence they were passaged by trypsinization, and the dose of 5-FU was increased.
7
Regulation of MALAT1, miR-206, ARNT, and PPARα in HepG2 cells
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The shRNA MALAT1 (sh-MALAT1), sh-negative control (sh-NC), miR-206 inhibitor, inhibitor NC, miR-206 mimics, miR-NC, sh-ARNT and sh-PPARα were obtained from GenePharma (Shanghai, China). HepG2 cells transfected with sh-MALAT1, sh-NC, miR-206 inhibitor, inhibitor NC, miR-206 mimics, miR-NC, sh-ARNT or sh-PPARα by using Lipofectamine 3000 (Invitrogen).
8
Knockdown of GJA1 in EPCs
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Short hairpin RNA targeting GJA1 (sh-GJA1) and negative control (sh-NC) were purchased from RiboBio (Guangzhou, China). miR-206 mimics, miR-206 inhibitor, and the corresponding negative controls (miR-NC) were all synthesized by GenePharma (Shanghai, China). EPCs were inoculated into 6-well plates (2 × 105 cells/well), and transfection was performed using Lipofectamine 2000 (Invitrogen, CA, USA) when confluence reached 50%. After 48 h, the transfection efficiency was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR).
9
Lentiviral-Mediated Gene Manipulation
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Lentiviruses overexpressing FER, lentiviral-based small hairpin RNA (shRNA) against FER, miR-206 mimics and miR-206 inhibitor were purchased from GenePharma (Shanghai, China). Lentiviruses were used to infect HCC cells based on the manufacturer’s protocol. At 48 h after infection, the cells were incubated with puromycin (5 μg/mL) for two weeks to establish stable cell clones. The sequences are provided in Supporting Table 1
10
miR-206 Mimic and Inhibitor Assay
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The miR-206 mimics, negative control (NC) mimic, miR-206 inhibitors, and NC inhibitor were all purchased from GenePharma (GenePharma, Shanghai, China).
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