Fig. 4

LY6E deficiency inhibited mIL-1β production and foam cell formation induced by IFN-α and ICs. BMDMs (2 × 10.⁶) were electroporated with 300 nM LY6E siRNA (siLY6E) or control siRNA (siCtl) and then stimulated with or without IFN-α (100 U/ml), 10 μg/ml ICs, 100 ng/ml LPS, or a combination of these stimuli for 24 h. For inflammasome activation control, cells were stimulated with LPS (500 ng/mL) for 4 h, followed by treatment with nigericin (5 μM) for 1 h (LPS + Nig). The total cell lysates and supernatants were separately collected for the measurement of several proteins as indicated by Western blotting (A and C). The loading control for supernatants is shown with Ponceau S stain. Each data point represents one mouse, and values are fold changes relative to the mean of the siCtl group. The statistical results from several independent experiments are shown (B and D). BMDMs were stimulated with oxLDL in the presence or absence of IFN-α or ICs, and foam cell formation was measured by Oil Red O staining. The cells were examined via light microscopy, and the percentages of Oil Red O-positive cells in 5 microscopic fields for each independent experiment were determined and statistically analyzed (E, F, G and H). Moreover, BODIPY dye analysis was carried out according to the description in the Materials and Methods (I and J). Each data point represents one mouse, and the values are fold changes relative to the mean of the siCtl treatment, as determined by Western blotting. Statistical analysis was performed with two-way ANOVA with Holm‒Sidak multiple comparisons to compare differences among different treatments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001