Analysis of the interaction between A₁R and A_2AR proteins in living cells based on FRET imaging and batch processing method

The quantitative FRET analysis of living cells is a tedious and time-consuming task for freshman lacks technical training. In this study, FRET imaging and batch processing method were combined to analyze reagents-induced interactions of A(1)R and A(2A)R on cell membranes. Results showed that the method had taken less time than if cell-by-cell was analyzed. The accuracy and repeatability of FRET efficiency values were likewise improved by removing the interference from anthropogenic factors. Then this method was applied to rapidly analyze acetaldehyde-induced interactions, which analyzed hundreds of single-cell trends by one operation, and the results revealed that interactions were consistently attenuated in LX-2 cells, and statistical differences appeared after 30 min. Combined with batch processing method, procedures of cells FRET analysis have been greatly simplified without additional technical work, which has broad prospects in large-scale analysis of cellar protein interaction.