Figure 4
LRET robustly detects chemical inhibition of Rac1 activitation in 96-well plates. (a) 293 T cells expressing a LRET Rac1 biosensor (30 nm ER/K linker) alone (BS) or with Tiam1 (BS + Tiam1) were grown overnight in 96-well plates, then incubated with. Time-gated measurements were obtained following 4 h incubation with EHT 1864 (50 µM, 0.5% DMSO) and addition of a lysis buffer containing TMP-TTHA-cs124. (b) 293 T cells co-expressing the sensor and Tiam1 were grown in 96-well overnight with or without inhibitors in the media. Time-gated measurements were taken after adding the lysis buffer as in (a). Bar graphs depict mean FRET/donor (FRET/D) ratios measured for each condition. Error bars, sem (n ≥ 3 transfections/plates; 16 wells/plate for each condition).
