Figure 7:
(A) Synthesis of MAGL reactive probe 9. (B) Live cells were treated with probe 9 for 1 h, followed by 2 μM TCO-TAMRA for 30 min, cell lysis, and analysis by in-gel fluorescence (C) In-gel fluorescence signals for a dose response of probe 9. Probe 9 (32 nM, 1 h) was competed by pre-treatment with MAGL inhibitor KML29 (300 nM, 1 h). (D) KML29 also incorporates a HFIP carbamate warhead. (E) Dose response fitting of the fluorescence signals of MAGL normalized by the total protein amount indicated by Coomassie staining. Data are reported as mean ± SEM (n = 2). See Fig S-21 for Coomassie staining.