Mir-125b mimic, miR-125b inhibitor, and the miRNA NC (negative control) mimic and inhibitor were synthesized by RiBoBio (Guangzhou, China). The STAT3 and negative control siRNA were purchased from Sigma-Aldrich. MiRNA or siRNA were transfected into cells according to the manufacturer’s instructions using Oligofectamine™ as the transfection reagent (Life Technologies, CA, USA). Cells were harvested at 72 h after transfection and then used for experiments.
Mir 125b inhibitor
Manufactured by RiboBio
Sourced in China
The MiR-125b inhibitor is a laboratory product designed to inhibit the expression of the miR-125b microRNA. This product is intended for research use only and its core function is to provide a tool for studying the biological role of miR-125b in various cellular and molecular processes.
Automatically generated - may contain errors
Lab products found in correlation
Mir 125b mimic, by RiboBio (3 mentions)
Oligofectamine, by Thermo Fisher Scientific (1 mentions)
Control mirna, by RiboBio (1 mentions)
Stat3 sirna, by RiboBio (1 mentions)
E coli poly a polymerase, by New England Biolabs (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Sybr premix ex taq 2, by Takara Bio (1 mentions)
Rnaiso plus, by Takara Bio (1 mentions)
7500 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Primescripttm rt reagent kit with gdna eraser, by Takara Bio (1 mentions)
Control sirna, by RiboBio (1 mentions)
Nucleofector transfection kit, by Lonza (1 mentions)
Cancer cell isolation kit, by Panomics (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
3 protocols using mir 125b inhibitor
1
miRNA Transfection Protocol for STAT3
2
miR-125b Expression Analysis in Skin Conditions
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Total RNA was isolated from HaCaT cells, cholesteatoma, and the corresponding skin with RNAiso plus (TaKaRa Biotechnology Co. Ltd., Dalian, China). The integrity, purity, and concentration of total RNA were analysed by spectrophotometry and gel electrophoresis. Then, the RNA was polyadenylated using E. coli Poly (A) Polymerase (New England Biolabs, MA) and reverse-transcribed into cDNA with a PrimeScriptTM RT reagent Kit with gDNA Eraser (TaKaRa). Finally, quantitative real-time PCR was carried out using a SYBR®Premix Ex Taq™II (TaKaRa) and 7500 real-time PCR system (Applied Biosystems, USA). The relative expression of gene levels was calculated using the 2–ΔΔCT method. U6 served as an internal control normalised the expression data of miR-125b. The sequences of the primers: miR-125b-5p: 5’-TCCCTGAGACCCTAACTTGTGA-3’ miR-reverse: n5’-GCTGTCAACGATACGCTACG-3’; miR-RT primer: 5’-GCTGTCAACGATACGCTACGTAACGGCATGACAGTGTTTTTTTTTTTTTTTTTTTTTTT-3’; U6 forward: 5’-CTCGCTTCGGCAGCACA-3’ and reverse: 5’-AACGCTTCACGAATTTGCGT-3’; HaCaT cells were transiently transfected with miR-125b mimics, miR-125b inhibitor, control miRNA, STAT3 siRNA, and control siRNA (all from RiboBio Co. Ltd., Guangzhou, China), using Lipofectamine® 3000 reagent (Invitrogen, Carlsbad, CA, USA).
3
Isolation and Culture of NSCLC Cells
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NSCLC cells were isolated from surgical tumor tissues using Cancer Cell Isolation Kit (Panomics). According to the manufacture’s instructions, cells were cultured for 4 days at 37 °C under 5 % CO2 in complete RPMI 1640 medium (GIBCO, containing 10 % heat-inactivated fetal bovine serum supplemented with 2 mM glutamine, 100 IU/ml penicillin and 100 mg/ml streptomycin sulfate), and used for experimental research. MiR-125b mimics and miR-125b inhibitor were from Ribobio (Guangzhou, China). Human TP53INP1 expression plasmid was purchased from Origen. Human TP53INP1 siRNA was from Santa Cruz. The nucleofector transfection kit was purchased from Amaxa.
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