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. Author manuscript; available in PMC: 2012 Jul 29.
Published in final edited form as: Chem Biol. 2011 Jul 29;18(7):846–856. doi: 10.1016/j.chembiol.2011.05.009

Figure 2. Stable shRNA-mediated knockdown of MAGL lowers FFA levels in PC3 cells.

Figure 2

(A) MAGL was stably knocked down using two independent short-hairpin RNA (shRNA) oligonucleotides (shMAGL1 and shMAGL2), resulting in >75 % reduction in MAGL activity in PC3 cells, as assessed by ABPP analysis of PC3 soluble proteomes, compared to shControl cells expressing an shRNA that targets a distinct serine hydrolase (DPP4). (B) Total C20:4 MAG hydrolytic activity of parental, shControl, and shMAGL1 and 2 PC3 whole cell lysate proteomes show significantly reduced MAGL activity in shMAGL cancer cells. (C, D) shMAGL cells show elevations in MAGs (C) and reductions in FFAs (D). The MAGL activity and MAG and FFA levels of shControl cells did not differ significantly from those of parental cancer cell lines. (E) Lipidomic analysis of PC3 shMAGL versus shControl cells shows not only elevations in MAGs and reductions in FFAs, but also lower levels of lysophosphatidyl ethanolamines (LPEs), phosphatidic acids (PAs), and lysophosphatidic acid (LPA). *p<0.05, **p<0.01 for shMAGL versus shControl groups. Data are presented as means ± SEM; n=4–5/group. See also Figure S1.