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The siRNAs were dissolved in an in vivo transfection reagent (TransIT In Vivo Gene Delivery System, Mirus) under sterile conditions

Plasma was collected at timepoints 0, 15 and 60 min by tail vein blood sampling and insulin concentrations were measured by enzyme-linked immunosorbent assay (ELISA

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Subsequently, NanoSU6668 (ICG), MNS (ICG) and T-MNS (ICG) (25 M) were added to the culture medium and incubated for an additional 6 h

The cohort had median age 64 years (interquartile range [IQR] 58, 69), was 97.6% male, and 70.8% White

The kit used a unique molecular identifier (UMI) of 8 random bases to label pre-amplified cDNA molecules, thereby removing repetitive bias in PCR and sequencing steps