FLIM analysis of the effect of GSMs and HP on APP-CT positioning relative to the membrane . A) APP/APLP2 dKO cells co-transfected with myrGFP and wild-type APP-RFP were treated with vehicle control, fenofibrate (FF), celecoxib (cele), ibuprofen (ibu), indomethacin (indo), or flurbiprofen (flurbi). The donor lifetime shortened in cells treated with FF and cele, and increased in cells treated with ibu, indo and flurbi, reflecting increased and decreased proximity between the APP-CT and membrane, respectively, compared to that in cells treated with the DMSO vehicle. mean ± SD; *p < 0.05; **p < 0.001, ANOVA; n = 42-84 cells analyzed per condition in three independent experiments. B) PS1/2 dKO cells were co-transfected with myrGFP and APP-RFP, and treated with fenofibrate or ibuprofen. Neither ibuprofen nor fenofibrate treatment significantly changed the myrGFP lifetime in PS1/2 dKO cells. C) PS1/2 dKO cells (green bar) and PS1/2 dKO + wild-type PS1 cells (black bars) co-transfected with myrGFP and APP-RFP were treated with either DMSO or HP to prevent APP-PS1 interactions. Three to five independent experiments were performed. Data from a representative experiment is shown. n: number of cells analyzed in the experiment (*p < 0.01; **p < 0.05; n.s. - not significant; ANOVA).