Behavior of distinct mRuby3 and mScarlet containing tandem constructs.

(A) The lifetimes of a distinct set of NG-Stop express cells with the lifetimes of 2 new mRuby3 containing tandem constructs and an mScarlet containing construct. Black bars indicate the average ± 95% confidence interval. *** = P < 0.0005 compared to NG-Stop. NG-Stop and NG-P2A-mRuby3 are not statistically different (P = 0.65). N for the lifetime measurements of each construct is as follows NG-Stop: 64 cells, mRuby3-GGSGG-NG: 66 cells, NG-P2A-mRuby3: 69 cells, and NG-mScarlet: 77 cells. (B) Average FRET efficiency for each of the new tandem constructs. (C) Example merge confocal images of HEK293 cells expressing each of the new tandem constructs. Pooled histograms of the slopes of cells expressing (D) mRuby3-GGSGG-NG, (E) NG-P2A-mRuby3, and (F) NG-mScarlet. N for the confocal measurements of each construct is as follows mRuby3-GGSGG-NG: 2302 cells, NG-P2A-mRuby3: 1548 cells, and NG-mScarlet: 1150 cells.