Fig. 3

MiR-146a deficiency promotes synthesis and secretion of pro-inflammatory cytokines. Bone-marrow-derived macrophages (BMDMs) prepared from miR-146a knockout (KO) mice and wild-type (WT) B6 mice were incubated with endotoxin-free monosodium urate (MSU) crystals (0.25 mg/mL) for 0, 4, and 8 h. The IL-1β gene expression in BMDMs was detected via quantitative (q)PCR (a). IL-1β protein levels in BMDMs were measured by western blot (b). IL-1β protein levels were detected in BMDM culture medium (c) and in lavage fluid from the peritoneal cavity (d) using ELISA. TNF-α gene expression was measured by qRT-PCR in BMDMs (e). The ratio of BMDMs producing TNF-α was measured by fluroescence-activated cell sorting (FACS) (f). Values are the mean ± SEM of three to five individual experiments using cells from three different mice of each genotype; groups were compared using the t test