(A) Glycerol-3-phosphate dependence of GPAT was determined by incubating 10–20 μg total particulate in the presence of 82.5 μM 16:0 CoA and varying the concentration of G3P from 10 to 800 μM, in the absence or presence of NEM. All experiments were performed in triplicates. Data are presented as mean ± SEM. See also Figure S6.
(B) The recovery of differentiation (day 8) is shown by Oil red O staining of WT, Bscl2 KO, and Bscl2 KO MEF cells treated with 40 μM inhibitor during the first 2 days of differentiation.
(C) Total RNA extracted from wild-type and Seipin−/− preadipocytes cultured in the presence or absence of GPAT inhibitor was used to measure the indicated mRNA levels by qPCR. Values represent mean ± SD of three independent experiments performed in triplicate. Statistical analysis was performed using Student’s t test. *p < 0.05, inhibitor treatment versus DMSO in Seipin KO cells.