Fig. 2.

Packaging and delivery of RNAs via ARMMs. a RNA packaging strategy. TAT peptide, which binds specifically to TAR, is fused to the C-terminus of ARRDC1 to recruit RNA cargo molecules linked to TAR, into ARMMs. b Packaging of TAR-GFP mRNA in ARMMs. ARRDC1-TAT was co-transfected with TAR-GFP or control GFP construct into HEK293T cells. ARMMs were pelleted via ultracentrifugation. qRT-PCR was done on ARMMs and on the transfected cells for GFP and for a control mRNA (HPRT1). c Packaging of TAR-p53 mRNA in ARMMs. TAR-p53 was co-transfected with ARRDC1 or ARRDC1-TAT construct into HEK293T cells. ARMMs were pelleted via ultracentrifugation. qRT-PCR was done on ARMMs and on the transfected cells for TAR-p53 and for HPRT1. d Transfer of TAR-GFP mRNA into recipient cells. A549 cells were incubated with ARMMs containing TAR-GFP mRNA overnight, washed with PBS extensively, and subjected to mRNA analysis by qRT-PCR. e Transfer of TAR-p53 mRNA into recipient cells. p53-null H1299 cells were incubated with ARMMs containing TAR-p53 mRNA overnight, washed with PBS extensively, and subjected to mRNA analysis by qRT-PCR. f Translation of ARMMs-delivered GFP mRNA in recipient cells. A549 cells were incubated with ARMMs containing TAR-GFP mRNA for 24 h with or without the translational inhibitor cycloheximide (CHX), and subjected to flow cytometry analysis. g Activation of p53 target genes in recipient cells receiving TAR-p53 ARMMs. P53-null H1299 cells were incubated with ARMMs containing TAR-p53 mRNA for 18 h and subjected to mRNA analysis by qRT-PCR to detect MDM2 and p21 mRNAs. At least three independent replicates were done for all assays. Data were presented as the mean ± SD in the bar graphs. *p < 0.05; **p < 0.01