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Platelet-derived exosomes as the nano-carrier for miR-150 to modulate gene expression and cell cycle in the M07-e cell line 1 1 Introduction 1 2 Materials and methods 2 2.1 Exosome isolation and characterization 2 2.1.1 Dynamic light scatter 2 2.1.2 Transmission electron microscopy (TEM) 2 2.1.3 Western blot 2 2.2 Exosome loading with miR-150 2 2.2.1 Cell culture 3 2.3 Cell transfection with exosomes 4 2.4 Molecular study 4 2.4.1 Total RNA extraction 4 2.4.2 cDNA synthesis and RT-PCR miR-150 4 2.4.3 cDNA synthesis and RT-PCR c-myb 4 2.5 Cell cycle analysis by PI staining 5 2.6 Statistical analysis 5 3 Results 5 3.1 Characterization of exosomes 5 3.2 Efficient loading of miR-150 into exosomes 5 3.3 Cellular uptake of loaded exosomes 5 3.4 Molecular study 5 3.4.1 RT-PCR confirmed the successful transfer of miR-150 into the megakaryocytic cells via PLT-EXOs 5 3.4.2 c-myb mRNA expression reduced after miR-150 enhancement 5 3.5 Cell cycle analysis 5 3.5.1 Frequency of cells in the G2 phase of the TPO group increased 5 4 Discussion 6 5 Conclusion 8 Funding 8 Authors contribution 8 Declaration of competing interest 8 Data availability 8 Acknowledgements 8 References 8

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Language

EN

Category

nonfiction