Figure 3: SCIMP is required for a subset of proinflammatory cytokine responses in macrophages.

(a) SCIMP was silenced by siRNA in BMM. Representative immunoblots showing levels of SCIMP, IκB, phospho-JNK, phospho-p38 and phospho-ERK1/2 at 0, 30 and 60 min post-LPS stimulation. (b) Relative chemiluminescence of IκB, phospho-JNK, phospho-ERK1/2 and phospho-p38 was assessed at 30 min and 60 min post-LPS stimulation in SCIMP-silenced BMM. Graphs represent pooled data from n=3 experiments (mean+s.e.m.). (c,d) SCIMP silencing in BMM reduces proinflammatory cytokine production at the mRNA (c) and protein (d) level. Levels of individual mRNAs, relative to Hprt, at 4 h post-LPS stimulation were assessed by qPCR (n=4 experiments), and levels of secreted cytokines at 24 h post-LPS stimulation were assessed by enzyme-linked immunosorbent assay (ELISA; n=4 experiments). Graphs depict mean+s.e.m. (e) IL-6 and IL-12p40 protein levels were assessed by ELISA in SCIMP-silenced BMM treated with LPS or TNF for 24 h. Data is representative of two independent experiments. Graphs depict mean+range from technical repeats (n=2). Data in (a–d) are representative of, or combined from, at least 3 independent experiments. For (b–d), significance was assessed using one-way ANOVA (*P<0.05 and **P<0.01; NS, not significant).