Figure 4. The effect of lipin-2 on ASC oligomerization and inflammasome assembly. BMDMs from WT and Lpin2−/− mice (A–D) were treated with 200 ng/ml LPS for 4 h, 2 mM ATP for 40 min, or both, as indicated. (A) Proteins from whole-cell lysates and purified cross-linked ASC oligomers were analyzed by immunoblot using specific antibodies against ASC or β-actin as a loading control. (B and C) Cells were stimulated as indicated and stained with specific antibodies against ASC and DAPI, as mentioned in the Materials and methods. Fluorescence was imaged by confocal microscopy. Bars: (B) 10 µm; (C) 50 µm. Arrowheads denote ASC specks. (D) Fold increase in ASC speck production from 700–1,400 cells is shown. Data from A–D are representative of at least three independent experiments. Data from D are shown as means ± SD *, P < 0.05, Student’s t test.
Figure 4.

The effect of lipin-2 on ASC oligomerization and inflammasome assembly. BMDMs from WT and Lpin2−/− mice (A–D) were treated with 200 ng/ml LPS for 4 h, 2 mM ATP for 40 min, or both, as indicated. (A) Proteins from whole-cell lysates and purified cross-linked ASC oligomers were analyzed by immunoblot using specific antibodies against ASC or β-actin as a loading control. (B and C) Cells were stimulated as indicated and stained with specific antibodies against ASC and DAPI, as mentioned in the Materials and methods. Fluorescence was imaged by confocal microscopy. Bars: (B) 10 µm; (C) 50 µm. Arrowheads denote ASC specks. (D) Fold increase in ASC speck production from 700–1,400 cells is shown. Data from A–D are representative of at least three independent experiments. Data from D are shown as means ± SD *, P < 0.05, Student’s t test.

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