Fig 4.

Presentation of MERS-CoV RBD-mRNA and expression of the target protein in cells. (A) Presentation of mRNA in 293T cells. (a) Cy5-labeled mCherry-RBD-mRNA (1 μg/ml) was incubated with 293T cells at 37°C. The cells were collected at different times postincubation and analyzed for Cy5-positive cells by flow cytometry. (b) Representative images by confocal microscopy at 27 hours postincubation, with nuclei in blue, cell membrane (pan-Cadherin) in green, and Cy5-mRNA in violet. (c) Schematic images showing the time period of Cy5-labeled mRNA in 293T cells. (B) Detection of expression of mRNA-encoding protein in target cells. (a) Cy5-labeled mCherry-RBD-mRNA (1 μg/ml) was incubated with 293T cells at 37°C. The cells were collected at different times postincubation and detected for mCherry fluorescence (mRNA expression) by flow cytometry. (b) Representative confocal microscopy images of the expression of mRNA-encoding protein. mCherry is in red, lyso (lysosome) is in green, and nuclei are in blue. Empty LNPs were included as a control. (c) Schematic images showing the time period of mRNA-encoding protein expression in 293T cells. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)