Fig. 3

LY6E deficiency inhibited IL-1β expression induced by stimulation with 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 various stimuli as indicated for 24 h. The concentrations of the individual stimuli used were as follows: the TLR7/8 agonist R848 (2.5 μg/ml), the TLR3 agonist PolyIC (10 μg/ml), LPS (100 ng/ml), IFN-α (100 U/ml), and ICs (10 μg/ml). The mRNA and protein levels of LY6E were determined by qPCR (A) and Western blotting (B), respectively. The mRNA expression of several inflammation-associated molecules induced by various stimuli with or without LY6E deficiency conditions was determined (C). The IFN-α- and IC-induced expression of IL-1β with or without LY6E knockdown was determined by Western blotting (D and E). Each data point represents one mouse, and the values are fold changes relative to the mean of the siCtl in RT‒qPCR and Western blotting. For Western blotting, the samples were derived from the same experiment, and both the gels and the blots were processed in parallel. Statistical analysis was performed with Student’s t test to compare the means between two groups (A and B) or two-way ANOVA with Holm‒Sidak’s multiple comparisons test to compare differences among different treatments (D and E). *P < 0.05, **P < 0.01, ****P < 0.0001