miRNA mimics, inhibitors, siRNAs (Genepharma, Shanghai, China) and plasmids transfections were performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol. The miR-133b mimics, miR-133b inhibitor, NEAT1 siRNA (si-NEAT1), TIMM17A siRNA (si-MAML) and their negative controls were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China). The sequences of miR-133b mimics, miR-133b inhibitor, si-TIMM17A and their negative controls were list in the Supplementary Table S3 . Lentivirus (Genepharma, Shanghai, China) encoding miR-133b or TIMM17A were imported into MDA-MB-231 cells as previously described [38 (link)]. The clones with the stable miR-133b or TIMM17A expression were selected by green fluorescence protein (GFP) expression.
Mir 133b inhibitor
Manufactured by GenePharma
Sourced in China
The MiR-133b inhibitor is a laboratory reagent used to inhibit the expression of the microRNA miR-133b. MiR-133b is a small non-coding RNA molecule involved in the regulation of gene expression. The MiR-133b inhibitor can be used in research applications to study the biological functions and regulatory mechanisms of miR-133b.
Automatically generated - may contain errors
Lab products found in correlation
Mir 133b mimic, by GenePharma (3 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Mirna mimic, by GenePharma (1 mentions)
Lentivirus, by GenePharma (1 mentions)
Sirnas, by GenePharma (1 mentions)
Opti mem medium, by Thermo Fisher Scientific (1 mentions)
Inhibitor nc, by GenePharma (1 mentions)
Mimic nc, by GenePharma (1 mentions)
Lipofectamin 2000, by Thermo Fisher Scientific (1 mentions)
Oe nc, by Sangon (1 mentions)
Kyse 150, by Thermo Fisher Scientific (1 mentions)
Kyse 150, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Kyse30, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
Het 1a, by American Type Culture Collection (1 mentions)
Ec9706, by American Type Culture Collection (1 mentions)
Kyse450, by American Type Culture Collection (1 mentions)
Kyse30, by American Type Culture Collection (1 mentions)
Ec109, by American Type Culture Collection (1 mentions)
4 protocols using mir 133b inhibitor
1
Regulation of miR-133b and TIMM17A in Breast Cancer
2
Overexpression of COL1A1 via Plasmid Transfection
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NC mimic, miR-133b mimic, NC inhibitor and miR-133b inhibitor were obtained from GenePharma (Shanghai, China). The sequences of oe-NC (negative control vector), oe-COL1A1(COL1A1 overexpression vector) were synthesized by Sangon Biotech (Shanghai) Co., Ltd. The recombinant plasmid pEGFP1-COL1A1 was constructed using the pEGFP1 overexpression vector. Cells in 3×105 cells/well were seeded into a six-well plate and transfected when the growth density reached 50% according to the instructions of lipofectamin2000 (11,668–019, Invitrogen, California, USA). Four micrograms of recombinant plasmids and 10 μL lipofectamin2000 were diluted by 250 μL Opti-MEM medium (51,985,042, Gibco, Gaithersburg, MD, USA) without serum, respectively. After keeping at room temperature for 5 min, the two liquids were mixed together uniformly and added dropwise to the cell culture wells 20 min later. After shaking, cells were cultured in a cell incubator with 5% CO2 at 37 °C. Six h later, the medium was replaced and cells were harvested 48 h after transfection.
3
Regulation of ESCC Cell Lines by KCNQ1OT1 and miR-133b
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Human ESCC cell lines (KYSE150, KYSE30, KYSE450, EC9706, and EC109 cells) and human esophageal epithelial cells (Het-1A) were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific, Shanghai, China) containing 10% fetal bovine serum (FBS, Sciencell Research, Carlsbad, CA, USA) at 37°C in 5% CO2.
pcDNA empty vector (normal control, NC), pcDNA-KCNQ1OT1 (KCNQ1OT1), scrambled siRNA (normal control, si-NC), small interfering RNAs against KCNQ1OT1 (si-KCNQ1OT1#1 and si-KCNQ1OT1#2), miRNA control (mimics NC), miR-133b mimics, an inhibitor control (inhibitor NC), and miR-133b inhibitors were obtained from GenePharma Co., Ltd. (Shanghai, China). The KYSE30 and KYSE150 cells were transfected using LipofectamineTM 2000 (Invitrogen, Carlsbad, CA, USA). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the transfection efficiency after 24h of transfection.
pcDNA empty vector (normal control, NC), pcDNA-KCNQ1OT1 (KCNQ1OT1), scrambled siRNA (normal control, si-NC), small interfering RNAs against KCNQ1OT1 (si-KCNQ1OT1#1 and si-KCNQ1OT1#2), miRNA control (mimics NC), miR-133b mimics, an inhibitor control (inhibitor NC), and miR-133b inhibitors were obtained from GenePharma Co., Ltd. (Shanghai, China). The KYSE30 and KYSE150 cells were transfected using LipofectamineTM 2000 (Invitrogen, Carlsbad, CA, USA). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the transfection efficiency after 24h of transfection.
4
Modulating FOXL2 Expression via miR-133b Mimics and Inhibitors
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MiR‐133b mimics, miR‐133b inhibitors, and the related negative control (NC) were purchased from GenePharma. Two predesigned small interfering RNA (siRNA) sequences of FOXL2 were synthesized by GenePharma. The siRNA constructs were as follows: si‐FOXL2‐1: 5′‐GCUACCGCAGCCUCCCUCATT‐3′; si‐FOXL2‐2: 5′‐GCGUAGUGAACUCGUACAATT‐3′. We transfected siRNA into cells with Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer's instructions.
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