mRNA Sequencing
To detect potential changes in host cell gene expression patterns after transfection, parallel cultures of PC-3, Jurkat, and primary T cells were passaged into fresh plates or flasks. Control (untransfected) cultures were left to grow for 48 hours, while the remaining cultures were transfected with pEF-GFP one day after passaging. On the following day, a small fraction (10%) of each culture was then analyzed using flow cytometry and fluorescent microscopy to confirm EGFP expression, then total RNA samples were extracted from the remaining cells with a Qiagen RNEasy kit. The RNA samples were then submitted to the Beijing Genomics Institute (BGI, Hong Kong, China) for library preparation, next-generation sequencing, and data analysis to obtain the FPKM values shown in Tables 1-3.