Figure 5
From: A bimolecular fluorescence complementation flow cytometry screen for membrane protein interactions
Quantification of BiFC yields using flow cytometry. (A) Analysis of constructs compared in this study together with the single plasmid negative control (YFPC − CaM). The histograms show the distribution of fluorescence intensities in the log phase of growth. The fluorescence signal was evaluated from a threshold value of 500 intensity units, discriminating the YFP-signal from auto fluorescence. (B) Average fluorescence intensity is shown for each complex together with the standard error of the mean (n ≥ 10 technical repeats based on at least three biological repeats, i.e. independent transformation events, see Table 1). (C) The average of the fraction of fluorescent cells in percentage is shown for each complex together with standard error of the mean (n ≥ 10 technical repeats based on at least three biological repeats, i.e. independent transformation events). An unpaired two component t-test with Welch’s correction was used to determine statistical difference of fluorescence frequency between YFPN − AQP0 + YFPC − CaM and YFPN − AQP0∆ + YFPC − CaM as well as YFPN − AQP0 + YFPC − AQP0 and YFPN − AQP0∆ + YFPC − CaM with p values < 0.0001 demonstrating a statistically robust discrimination above a certain yield, as indicated ****.
