Fig. 5
LY6E knockdown inhibited the IFN-α- and IC-induced expression of active caspase 1. BMDMs (2 × 10.6) were electroporated with 300 nM siLY6E or siCtl and then stimulated with or without IFN-α or ICs for 24 h. As a positive control, LPS (500 ng/mL) was used to stimulate the cells for 4 h, followed by treatment with nigericin (5 μM) for 1 h (LPS + Nig). The expression of active caspase 1 was determined by flow cytometry in lysates (A, B, and C) or by Western blotting in supernatants (D, E, and F). NLRP3 expression was measured by Western blotting (G and H). The correlation between LY6E and NLRP3 mRNA levels in monocytes from SLE patients was determined (I). In parallel, the expression of complement 5a receptor 1 (C5ar1) and C5ar2 mRNAs was determined by qPCR (J). Each data point represents one mouse, and the values are fold changes relative to the mean value of the siCtl in RT‒qPCR and Western blotting. The values for the flow cytometry results are presented as the geomeans (MFIs). Two-way ANOVA with Holm‒Sidak multiple comparisons was used to compare differences among different treatments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001