Figure 8. Osteopontin enhances EGFR1 signaling via a mechanism that requires α5β1, RCP, and association of EGFR1 with RCP. Cells were transfected with short hairpin vectors targeting β3, α5, β1, RCP or EGFR1 or with truncated GFP-RCP379-649 as indicated. Cells were serum starved overnight followed by a 40-min treatment with either 1 μM cilengitide or 0.5 μg/ml osteopontin. The cells were then challenged with EGF for the indicated times and lysed. Levels of active phosphoTyr845-EGFR1, phosphoSer473-Akt, and phosphoThr202/phosphoTyr204-ERK were determined by Western blotting. Protein loading of β-actin, Akt1, and EGFR1 was confirmed by Western blotting.
Figure 8.

Osteopontin enhances EGFR1 signaling via a mechanism that requires α5β1, RCP, and association of EGFR1 with RCP. Cells were transfected with short hairpin vectors targeting β3, α5, β1, RCP or EGFR1 or with truncated GFP-RCP379-649 as indicated. Cells were serum starved overnight followed by a 40-min treatment with either 1 μM cilengitide or 0.5 μg/ml osteopontin. The cells were then challenged with EGF for the indicated times and lysed. Levels of active phosphoTyr845-EGFR1, phosphoSer473-Akt, and phosphoThr202/phosphoTyr204-ERK were determined by Western blotting. Protein loading of β-actin, Akt1, and EGFR1 was confirmed by Western blotting.

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