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. 2010 Oct;88(4):769–778. doi: 10.1189/jlb.0410220

Figure 4. sLRP1 initiates cell signaling in macrophages.

Figure 4.

(A) RAW 264.7 cells were cultured in SFM for 1 h and then challenged with 30 nM sLRP1 for the indicated times. Immunoblot analysis was performed to detect the indicated signaling proteins. P-, Phosphorylated. (B) RAW 264.7 cells were treated with sLRP1 (50 nM), boiled sLRP1 (50 nM), or vehicle for 20 min. Phosphorylated and total p38 MAPK were determined by immunoblot analysis. (C) RAW 264.7 cells were treated with increasing concentrations of sLRP1 for 20 min. Phosphorylated and total p38 MAPK were determined. Immunoblots were subjected to densitometry. The results of 3 experiments were averaged (mean±sem; *P<0.05). (D) RAW 264.7 cells in which LRP1 was stably silenced and control cells that were transfected with empty vector (pSUPER) were cultured in SFM for 1 h and then treated with sLRP1 (50 nM), TNF-α (10 μg/ml), or vehicle for 20 min. Phopho-p38 MAPK and total p38 MAPK were determined by immunoblot analysis. (E) Primary cultures of mouse BMMs were isolated from mice in which LRP1 was conditionally deleted in macrophages (LRP1−/−) and from WT mice in the same genetic background. These cells were treated with sLRP1 (50 nM), LPS (10 ng/ml), or vehicle for 20 min. Immunoblot analysis was performed to detect phospho-p38 and total p38.