The BMSCs were transfected with negative control (NC) mimics or miR-216a-5p mimics (Genepharma, China) using Lipofectamine 3000 (Invitrogen, United States) per the kit’s procedures. After 72 h of transfection, BMSCs were cultured with medium containing exosome-free FBS (Gibco, United States) for an additional 24 h. Afterwards, the exosomes of transfected BMSCs were isolated (named BMSC-NC-Exo and BMSC-miR-Exo), and reverse transcription quantitative polymerase chain reaction (RT-qPCR) was completed for detecting the level of miR-216a-5p in BMSCs or BMSC-Exo.
Mir 216a 5p mimics
Manufactured by GenePharma
Sourced in China
MiR-216a-5p mimics are synthetic RNA molecules designed to mimic the function of the naturally occurring microRNA, miR-216a-5p. MicroRNAs are small, non-coding RNA molecules that play a crucial role in regulating gene expression.
Automatically generated - may contain errors
3 protocols using mir 216a 5p mimics
1
Transfection and Exosome Isolation from BMSCs
2
DANCR Silencing in Chondrocytes
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The specific interference sequence for DANCR (si-DANCR) was constructed at Genechem (Shanghai, China), and then transfected into chondrocytes. The sequences were showing as follows: si-DANCR-1: 5′-UCGGAGGUGGAUUCUGUUA-3′, si-DANCR-2: 5′-AGCCAACTATCCCTTCAGT-3′. MiR-216a-5p mimics, miR-216a-5p inhibitors, and respective negative control miRNA were synthesized by GenePharma (Shanghai, China). Transfection was performed using Lipofectamine 3000 (Invitrogen, U.S.A.) according to the manufacturer’s instructions.
3
Bcl-2 Silencing via miR-216a-5p
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The siRNAs, miR-216a-5p mimics, inhibitors, and their respective negative control (NC) vectors were purchased from GenePharma (Shanghai, China) and then transiently and stably transfected according to the manufacturer’s instructions. The most effective interference sequences within Bcl-2 were identified based on real-time quantitative PCR (RT-qPCR) and then used for subsequent in vitro experiments. The stable overexpression or silence miR-216a-5p expression cell lines and control cell lines were constructed by lentivirus infection. The detailed procedure is provided in the “Supplementary materials” section. The relevant mimic and inhibitor sequences are listed in the “Supplementary materials” section.
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