Fig. 9

LY6E mediated its effects through regulating CMPK2. BMDMs were electroporated with siLY6E or siCtl and then stimulated with or without IFN-α, ICs, or LPS + nigericin (LPS + Nig) for 24 h, after which the Cmpk2 mRNA and protein levels were determined (A and B). BMDMs were electroporated with 300 nM siCMPK2 or siCtl and then stimulated with and without IFN-α or ICs for 24 h, and the levels of CMPK2 and LY6E in total cell lysates and both active caspase 1 (p20) and mIL-1β in supernatants were measured by Western blotting (C and D). BMDMs electroporated with siLY6E or siCtl were transduced with lentivirus carrying wild-type CMPK2-DYK or the control DYK vector. After 48 h, the medium was replaced with fresh medium, and the cells were then stimulated with IFN-α or ICs for 24 h. The measurement of mIL-1β in the supernatants (E and F) and mtDNA release into the cytosol (G and H) were carried out accordingly as previously described. 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 and ****, P < 0.0001