Integrated transcriptomic and metabolomic analysis reveals the effects of EMMPRIN on nucleotide metabolism and 1C metabolism in AS mouse BMDMs
Six RNA samples from the CON and siE groups of mouse BMDMs were sent for sequencing at BioTree (BioTree, Shanghai, China). Total RNA was extracted using the TRIzol reagent (Thermo Fisher, 15596018) according to the manufacturer's protocol. The quantity and purity of the total RNA were analyzed using the Bioanalyzer 2100 and the RNA 6000 Nano LabChip Kit (Agilent, CA, United States). Subsequently, mRNA was purified from the total RNA (5 ug) using Dynabeads Oligo (dT) (Thermo Fisher, CA, United States) with two rounds of purification. Following purification, the mRNA was fragmented into short fragments using divalent cations. The fragments were then reverse-transcribed into cDNA using random primers. The cDNA underwent a series of synthesis, modification, conversion, and PCR amplification processes to form libraries with an average fragment size of 300 bp +/- 50 bp (strand-specific libraries). Finally, 2 x 150 bp paired-end sequencing (PE150) was performed on an Illumina Novaseq 6000, following the vendor's recommended protocol. Gene expression abundance and variations were analyzed by calculating the FPKM (fragments per kilobase of transcript per million mapped reads) values using the StringTie software.