Figure 4: Measuring intracellular target engagement at bromodomain BRD4 using BRET.
From: Target engagement and drug residence time can be observed in living cells with BRET
(a) Specific and dose-dependent BRET was observed with Nluc-fusions to full-length BRD4 expressed in HeLa. BRET values at each tracer concentration were background-corrected by parallel measurements made in the presence of an excess of unmodified compound (20 μM) (described in Methods). Data are mean±s.d. of four data points. (b) Use of BRET to measure rank-order affinity of BRD inhibitor panel against BRD4 within live HeLa cells (1 μM IBET-NCT held constant). Data are mean±s.e.m. of four independent experiments.
