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. Author manuscript; available in PMC: 2012 Oct 1.
Published in final edited form as: Hepatology. 2011 Jul 29;54(4):1237–1248. doi: 10.1002/hep.24504

Figure 4. Cell-to-cell transfer of firefly luciferase by exosomes.

Figure 4

(A) Real-tine PCR was performed on an cDNA transcribed from an equivalent amount (600 ng) of RNA from PLC-luc derived exosomes or their donor cells (n = 3, each in duplicate). PCR amplification curves for Fluc mRNA and 18S rRNA are shown. (B) PLC/PRF/5 cells were incubated with 15 μg/ml of PLC-luc derived exosomes for 16 hours. RNA was isolated and equivalent amount of RNA (300 ng) was transcribed to cDNA (n = 3). Amplification curves by quantitative real-time PCR for Fluc mRNA and 18S rRNA in PLC/PRF/5 (recipient cells) and PLC-luc (donor cells) are shown. (C) PLC/PRF/5 cells in a 96-well plate were incubated with various concentrations of PLC-luc derived exosomes, and luciferase activity was assessed in these cells after 16 hours. Bars express the mean value of luminescence ± SEM of four separate determinations. *, p < 0.05. (D) PLC/PRF/5 cells were pretreated with 25 μg/ml of cycloheximide for 2 hours to inhibit de novo protein synthesis. Cells were washed with PBS and incubated with 100 μg/ml of PLC-luc derived exosomes. Luciferase activity was assessed in these cells after 6 hours. Bars express the mean value of luminescence ± SEM of five separate determinations. *, p < 0.05.