Figure 7.

Ficolin‐A (FCN‐A) promoted the M1 polarization of bone‐marrow‐derived macrophages (BMDMs) through a Toll‐like receptor 4 (TLR4)/MyD88‐dependent pathway in vitro. (a) The protein expressions of the purified GST‐FCN‐A and GST were determined by SDS–PAGE. (b) The extracted membrane proteins from RAW264.7 cells were incubated with the purified GST‐FCN‐A or GST proteins. Co‐immunoprecipitation analysis of the interaction between TLR4 of macrophage and GST‐FCN‐A was performed by using anti‐TLR4. Rabbit IgG was used as a negative control in co‐immunoprecipitation. (c, e) BMDMs, isolated from wild‐type (WT), TLR4^−/− or MyD88^−/− mice, were stimulated with FCN‐A (10 μg/ml) for 24 hr, then the expressions of iNOS and Arg‐1 from BMDMs were examined by Western blot analysis. (d, f) The levels of pro‐inflammatory cytokines IL‐1β in cell lysates, and secreted IL‐6, tumour necrosis factor‐α (TNF‐α) were detected by ELISA. (g, h) Western blot analysis of p‐IRAK1, p‐p65, p‐ERK1/2 and p‐JNK in the BMDM lysates of TLR4^−/−, MyD88^−/− or WT after stimulation with FCN‐A for 0–45 min. In d and f, values are mean ± SEM from three independent experiments. *P < 0·05, **P < 0·01.