Genome-wide identification of mRNAs involved in BCL2L2-PABNP1 regulation

RNA-Seq technology was used to explore mechanisms of BP on adipogenesis in cells transfected with adenoviruses expressing BP at a condition of MOI 300 and 48 h which was predetermined with fluorescence microscope and qPCR analysis . A total of 3,074 DEmRs were obtained by RNA-Seq analysis, among which 1,476 were upregulated and 1,598 were downregulated in cells overexpressing BCL2L2–PABPN1 compared with control groups . Overall design: The recombinant adenoviruses were constructed using the AdEasy system (Hanbio, Shang, China). BP CDS were inserted into the shuttle plasmid containing an enhanced green fluorescent protein (EGFP) and cyto-megalovirus promoters using Kpn ? and Xho ? sites, and homologous recombina-tion was performed in E. coli BJ5183 with adenoviral backbone pAdEasy 1. The recom-binant adenovirus plasmid was packaged in HEK-293A cells after linearized with Pac ?. Preadipocytes were infected with adenovirus virions at multiplicity of infection (MOI) of 300. At 48 h post-infection, cells were collected for RNA-Seq with Illumina NovoSeq 6000 platform (Illumina, San Diego, CA, USA) by Geneseeq Technology (Nanjing, China) using pair-end sequencing strategy according to the manufacturer's protocols. Cells treated with empty adenovirus were used as a control. A total of six Ribo-Zero RNA-sequencing libraries including overexpressing and control groups were con-structed, each with three replicates.