(A) L. salivarius total RNA (LAB RNA; 2 μg) was treated with or without RNase V1, then added to Raw264.7 cell culture in the presence or absence of FuGENE. IL-1β RNA was measured by qPCR. (B) LAB RNA was digested with indicated amounts of RNase III, RNase T1, RNase V1 or mock treated before adding to BMDM cell culture. 8 hr after incubation, total cell RNA was extracted to measure IL-1β expression by qPCR (upper panel). The efficiency of RNase treatment was verified by agarose gel electrophoresis (lower panel). (C) BMDM of the indicated genotypes was growing in the presence of LAB RNA at different concentrations for 8 hr, followed by the measurement of IL-1β RNA by qPCR. (D) BMDM of the indicated genotypes was growing in the presence of LAB RNA, R848 or LPS for 8 hr, then IL-1β induction was measured by qPCR. (E) Similar to (D), except that BMDM from Unc93b1 mutant mice (3d) was used. (F) BMDM from WT or TLR11−/− mice was incubated with LAB RNA or LPS followed by measurement of IL-1β RNA by qPCR. Error bars represent standard error of triplicate assays. N.D: not detected.