Metabolic Labeling and Inhibition Assay

17-ODYA labeling was performed as described previously ^{8 (link)}. HDFP, ML348, and ML349 syntheses were previously reported^{8 (link), 20 (link)}. Other inhibitors were purchased from Sigma (2-bromohexadecanoic acid, TVB-3166, and ABC44) or Cayman (JW480, WWL229, and WWL70). At ~90 % confluency, the growth media was aspirated and new media containing the inhibitor was added to the cells (Control = 0.1% v / v DMSO final). For ABPP and acyl-RAC proteomic experiments, cells were incubated with HDFP for 3 hours. After incubation, 20 µM 17-ODYA (Cayman) was added. At different time points, cells were washed with cold DPBS (Invitrogen) and scraped off the plate. The cell pellets were washed with DPBS and stored at −80 °C.