Figure 2
From: Defects in fatty acid amide hydrolase 2 in a male with neurologic and psychiatric symptoms
Functional analysis of FAAH2 mutations. (A) Expression of FLAG-tagged FAAH2 and FAAH2 mutants in 293T cells. The blots were probed with anti-FLAG (Sigma # F1804) and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH, Abcam #ab8245) antibodies. (B) AEA and PEA hydrolysis by cell homogenates expressing FAAH2 and FAAH2 mutants. Results were analyzed using one-way ANOVA followed by Dunnett’s post hoc analysis. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (n = 5–7). (C) Expression of FLAG-tagged FAAH1 and the FAAH1 p.Ala478Ser mutant in 293T cells. Blots were probed with an anti-FLAG antibody. (D) AEA hydrolysis by WT and p.Ala478Ser FAAH1 ***, p < 0.001 (n = 4). (E) Human fibroblasts do not express FAAH1. Fibroblasts from a control patient and the FAAH2 p.Ala458Ser affected patient were probed with anti-FAAH1 antibodies (Abcam #ab54615). FAAH1 transfected and untransfected 293T cells served as controls. (F) Expression of FAAH2 in fibroblasts from the affected p.Ala458Ser patient and an unaffected control. Blots were probed with an anti-FAAH2 antibody (Abcam, ab103724). (G) AEA hydrolysis by control fibroblasts and affected FAAH2 p.Ala458Ser fibroblasts in the absence and presence of URB597. *, p < 0.05; **, p < 0.01 versus unaffected control. #, p < 0.05 versus affected control (n = 4).