Q5000 ultramicro spectrophotometer

The total RNA of mouse mammary gland tissue and MAC-T cells was extracted with TRIzol reagent (Solarbio, Beijing). Q5000 ultramicro spectrophotometer (Quawell, USA) was used to evaluate RNA purity and measure RNA concentration. Hifair® III 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) and Hieff® qPCR SYBR® Green Master Mix were acquired from Yeasen (Shanghai, China) and used to examine target gene expression by qPCR with 2^−ΔΔCq method. The primers used in this study are shown in Table 1.

The total RNA of mouse hippocampus was extracted by homogenization of Trizol on ice. After measuring and evaluating the purity and quality by Q5000 Ultramicro spectrophotometer (Quawell, Los Angeles, CA, USA), high-throughput sequencing of mRNA and miRNAs was performed, and the original data of sequencing was determined according to the difference. The upregulation of miRNAs regulates differentially downregulated mRNA and differentially downregulates miRNAs targeting the upregulation of mRNA, and differential miRNAs and differentially expressed mRNA. It also performs functional analysis of GO and KEGG associations of miRNAs targeting regulatory gene sets. The method was reported in Figure 1.