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. 2018 Jan 29;115(7):E1530–E1539. doi: 10.1073/pnas.1716095115

Fig. 2.

Fig. 2.

OLT1177 inhibits NLRP3 inflammasome formation. (A) Immunoprecipitation (IP, Left) and analysis (Right, mean of 3 ± SEM) of the NLRP3-ASC association in J774A.1 cells stimulated with LPS/nigericin (NIG). (B) ATPase activity of recombinant NLRP3 in the presence of OLT1177 (n = 3), Bay11-7082 (BAY) (n = 2), and 3,4-methylenedioxy-β-nitrostyrene (MNS) (n = 2). (C–E) Immunofluorescent staining and (F–H) FRET from NLRP3 (in red) and ASC (in green) in J774A.1 cells stimulated with LPS and NIG in presence of OLT1177 (10 µM). Representative ASC speck-like structures are indicated (white arrows) and highlighted in the Inset (D). Images were acquired using Slide Book 6 (Intelligent Imaging Innovation). (I and J) Mean ± SEM of FRET intensity from NLRP3-ASC (I) and percent change of FRET intensity from NLRP3-caspase-1 (J). ***P < 0.001, **P < 0.01, *P < 0.05. The data are mean of three independent experiments. a.l.u.f.i., arbitrary linear units of fluorescence intensity.