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. 2013 Jun 18;8(6):e67170. doi: 10.1371/journal.pone.0067170

Figure 9. Two-color flow cytometry to quantitate fraction of cells that carried reporter plasmids.

Figure 9

(A) Negative control in the absence of both plasmids, which was used to gate cell populations in transfected cells. (B) 293T cells transfected with 0.6 μg pEGFP-Vpr and 0.2 μg pmCherry-Vpr in 2-ml volume in a 35-mm dish. The gates established based on negative control were used to quantitate fractions of cells that carry EGFP only, mCherry only and both fluorescent proteins in a mixed populations, with the percentages of each population indicated in the cytogram. (C) 293T cells transfected with 0.6 μg pEGFP-Vpr and 0.2 μg pmCherry-Vpr, with a media change six hours post transfection; (D) 293T cells transfected with 0.2 μg pEGFP-Vpr and 0.6 μg pmCherry-Vpr; (E) 293T cells transfected with 0.2 μg pEGFP-Vpr and 0.6 μg pmCherry-Vpr, with a media change six hours post transfection. All cell samples were collected at 48 hours post transfection. 488 nm and 560 nm lasers were used for excitation of EGFP and mCherry respectively. Two independent analyses yielded results that were identical within errors.