ECs were collected and total RNA was obtained with mirVANA miRNA kit. The RNA was transcribed using TaqMan® Advanced miRNA cDNA Synthesis Kit (Applied Biosystems; CA, EU) protocol. All assays were performed in duplicates for the differentially expressed miRNAs including miR-126-5p and -3p, let7g-5p, miR-103a-3p and miR-30b-5p (Life Technologies; CA, EU) (n = 5). For normalization miR-423-3p and let-7a-5p means were used as endogenous miRNA controls.26 (link) The amplification was performed in ABI PRISM 7900HT Sequence Detection System (Applied Biosystems; CA, EU) in 384-well plates. The amplification curves were analysed using the Applied Biosystems Sequence Detection System 2.4.1 software. Ct values were obtained and processed by the standard curve method and normalized with the endogen. To obtain fold difference vs. control the ratio sample/control was calculated. Networks, functional analyses, detection of miRNAs targets and interactions were generated by Ingenuity Pathway Analysis software (IPA) and RNA hybrid tool.
Let7g 5p
Manufactured by Thermo Fisher Scientific
Let7g-5p is a short, non-coding RNA molecule that functions as a microRNA. It is involved in the regulation of gene expression. This product is designed for research use only and its core function is to facilitate the study of microRNA biology.
Automatically generated - may contain errors
Lab products found in correlation
Sequence detection system 2, by Thermo Fisher Scientific (1 mentions)
Taqman advanced mirna cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Abi prism 7900ht sequence detection system, by Thermo Fisher Scientific (1 mentions)
Let 7g, by Thermo Fisher Scientific (1 mentions)
Control sirna, by Thermo Fisher Scientific (1 mentions)
2 protocols using let7g 5p
1
miRNA Expression Analysis Protocol
2
miRNA Regulation of Glucose Metabolism in 3T3-L1 Adipocytes
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3T3-L1 cells were transfected with mirVana miR-106b (LifeTechnology MC10067), let 7g-5p (LifeTechnology MC11758), 330-3p (LifeTechnology MC10732), 19b-5p (LifeTechnology MC13042), 142-3p (LifeTechnology M C10398),125a-5p (LifeTechnology MC12561), 331-3p (LifeTechnology MC10881) mimics or miR-106b or Let7g antagomirs (AM10067, MH11758 respectively). 72 h after transfection, 3T3-L1 cells were evaluated for 2-DG uptake and mRNA expression in lysates by RT-qPCR. 3T3-L1 cells were also transfected with pre-miR-106b siRNA sense oligonucleotides 5’-CCU AAU GAC CCU CAA GCC GUU-3 and antisense 5’-CGG CUU GAG GGU CAU UAG GUU-3’ then exposed to VD( + ) HSC or VD(-) HSC-recipient peritoneal macrophage media for 72 h before assessment of pre-miR-106-5p or mature miR-106b-5p. Peritoneal macrophages from VD( + ) HSC or VD(-) HSC recipients were transduced with lentivirus containing sense either Jarid2-siRNA (LifeTechnology 4390771), Ppargc1a-siRNA (LifeTechnology, AM16708), or control-siRNA (LifeTechnology AM461) for 48 h and mRNA expression was determined as previously described81 (link).
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