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. 2015 Jan 9;125(2):665–680. doi: 10.1172/JCI78253

Figure 6. FASN regulates NLRP3 and IL-1β expression through AKT activation in macrophages.

Figure 6

(A) Immunoblot analysis for NLRP3 and IL-1β and (B) measurement of TGs from wild-type BMDMs pretreated with C75 (0, 10, 20, and 50 μM) as well as (C) measurement of PEP, citrate, and lactate production from wild-type BMDMs pretreated with C75 (20 μM) for 2 hours before stimulation with LPS (500 ng/ml, 4 hours). β-Actin served as the standard. **P < 0.01, ANOVA. (D) Immunoblot analysis of AKT Ser473 phosphorylation in wild-type BMDMs pretreated with C75 (20 μM) for 2 hours before stimulation with LPS (500 ng/ml; 0, 10, and 20 minutes). Total AKT served as the standard. (E) Immunoblot analysis of AKT Ser473 phosphorylation in cell lysates from wild-type BMDMs pretreated with C75 (0, 10, 20, and 50 μM) for 2 hours before stimulation with LPS (500 ng/ml, 4 hours). Total AKT served as the standard. (F) Immunoblot analysis of AKT Ser473 phosphorylation in cell lysates from wild-type peritoneal macrophages transduced with lentiviruses expressing nontarget shRNA or 3 independent shRNA for FASN after stimulation with LPS (500 ng/ml, 4 hours). Total AKT served as the standard. (G) Immunoblot analysis for NLRP3, IL-1β, and AKT Ser473 phosphorylation in cell lysates from wild-type BMDMs pretreated with BX795 (10 μM) for 1 hour before stimulation with LPS (500 ng/ml, 4 hours). Total AKT served as the standard.