Figure 2.
Expression of reporter genes upon transfection into BHK cells of 500 ng of mRNA in non-self-amplifying (EYFP and Luc) and self-amplifying/replicon (Nod.EYFPOAS and Nod.LucOAS) form, using lipofectamine seeded into the 6-well cell culture plate (8 × 105 BHK-21 cells/well). (A) Expression of EYFP was captured with fluorescence microscopy; scale bar represents 400 μm. (B) Fluorescent cell population of EYFP levels from mRNA and replicon RNA was determined by fluorescence-activated cell sorting (FACS). The graphs are plotted with side scatter area [FSC-A] versus EYFP fluorescence intensity. For statistical significance, p value was calculated using unpaired two-tailed test. * p < 0.005. (C) Expression of Renilla luciferase for mRNA (Luc, EYFP) and replicon forms of the reporter gene, measured from the cell lysate collected post transfection. Multifold change of Renilla luciferase and EYFP mRNA levels for mRNA and replicon forms in transfected BHK-21 cells was determined by quantitative real-time PCR. The levels of respective mRNAs in the transfected cells were normalized by GAPDH mRNA.