CETSA Classics for in-cell AR target engagement. Representative western blots showing thermostable AR following indicated heat shocks in the presence (+) or absence (−) of (a) Enzalutamide (100 µM), (b) DHT (100 nM), or (c) DHT (100 nM) with Enzalutamide (30 µM). In each case a single cropped western blot is shown. Full-length blots are presented in Supplementary Fig. S11. (d) Quantification of thermostable AR western blots from CETSA Classics. Blot intensity (I) is normalised to intensity from the 40 °C sample. Thermal stabilisation was observed upon addition of the agonist dihydrotestosterone (DHT, 100 nM), but not by addition of the antagonist Enzalutamide (100 µM). Co-dosing Enzalutamide with DHT reversed DHT-induced thermal stabilisation in a concentration dependent manner. Data the mean ± SD of n ≥ 3, from ≥3 replicates.