TABLE 1.

Inhibitor profiles of hABHD6 and hABHD12

	hABHD6	hABHD12
Inhibitor	log[IC50] ± SEM	log[IC50] ± SEM
MAFP	−7.77 ± 0.04	−7.06 ± 0.04
IDFP	−6.97 ± 0.05	−5.60 ± 0.06
HDSF	−6.84 ± 0.10	−5.67 ± 0.11
THL	−7.32 ± 0.06	−6.72 ± 0.07
RHC-80267	−6.18 ± 0.08	Remaining activity, 96.7 ± 0.5% at 10 M
WWL70	−7.07 ± 0.05	Remaining activity, 101.0 ± 0.8% at 10 M
Pristimerin	−5.86 ± 0.07	Remaining activity, 101.4 ± 3.4% at 10 M

HEK293 cells were transiently transfected with the cDNAs encoding hABHD6 or hABHD12 as detailed in Materials and Methods. After 48 h, cells were harvested, and lysates were prepared for hydrolase activity measurements as described in Fig. 1. Cellular lysates (0.3 µg/well) were preincubated for 30 min at RT with DMSO (control) or with increasing concentrations of the inhibitors (MAFP, IDFP, HDSF, THL, RHC-8027, WWL70, and pristimerin). Thereafter, glycerol assay mix containing 2-AG (12.5 µM final concentration) was added, and glycerol production was monitored kinetically for 90 min at RT. Inhibitor dose-response curves were determined, and the IC₅₀ values were calculated as nonlinear regressions using GraphPad Prism 5.0 for Windows. Data are mean ± SEM from three independent experiments.