Application examples of the dual fluorescence RNA-binding assay. (A,B) Cells expressing eGFP, ACTB–eGFP, H2B–eGFP, MOV10–YFP, hnRNPC–eGFP, PABP–eGFP, FAM98A–eGFP, FAM32A–eGFP, or ENO1–YFP were induced with tetracycline for 16 h and irradiated with 254-nm UV light. After lysis, control samples were RNase-treated and recombinant proteins were immunoprecipited with GFP-multiTrap. Co-immunoprecipitated RNAs were hybridized with oligo(DT) coupled with one molecule of WellRED D4. The graphs represent relative green (eGFP/YFP) and far red (WellRED D4) fluorescence levels (A) and their ratios normalized to RNase-treated samples (B). The graphs were generated from three biological and three technical replicates (total of nine immunoprecipitations). (*) P < 0.05; (**) P < 0.01; (***) P < 0.001. Horizontal red and green lines represent averaged eGFP or Tred background signal, respectively, extracted from control samples.
