Figure 5

Inflammation and epidermal growth factor receptor analysis. (A) Sagittal sections of knees from wild-type (WT) and inactivated mitogen-inducible gene 6 (Mig-6^−/−) mice 4 weeks after surgery stained for monocytes, neutrophils and macrophages (brown). C: control knee and S: surgery knee. (B) Phosphorylated epidermal growth factor receptor (phospho-EGFR) staining (red) of knees from WT and Mig-6^−/−mice counterstained with 4′,6-diamidino-2-phenylindole (DAPI blue) (original magnification, 10×) 4 weeks after surgery. ImageJ software was used to quantify the area of phospho-EGFR staining (mean ± SE). (C) Total EGFR staining (red) of knees from WT and Mig-6^−/−mice counterstained with DAPI (blue) (original magnification, 10×) 4 weeks after surgery. ImageJ software was used to quantify the area of total EGFR staining (mean ± SE). (D) Western blots from control (C) and surgery (S) knees isolated from WT and Mig-6^−/−mice probed for phospho-EGFR, phosphorylated extracellular signal-regulated kinase (phospho-ERK), total ERK and β-tubulin. Densitometry (mean ± SE) was determined using ImageJ software. The positive (+C) and negative control (−C) lysates were protein extracts from calvarial osteoblasts isolated from 3-day-old WT mice subjected to either 60 minutes of 2 Pa of fluid shear stress or no mechanical stimulation. Gel lane numbers are written on some of the blots.