Quantitative real-time PCR analysis
Following the manufacturer’s instructions, total RNA was isolated from mouse liver using Trizol reagent (Invitrogen, USA). And then reverse-transcribed into cDNA using the qPCR RT kit(GeneCopoeia, USA),The cDNA was analyzed using real-time qPCR with a StepOnePlusTM Real-Time PCR System(Thermofisher, USA). Each reaction was performed in triplicate in a 96-well plate. The expression of each gene was normalized by the expression of GAPDH. miRNA and mRNA primers are listed in Table 1.
Table 1. The sequences of miRNA and mRNA primers