Figure 3. Emdogain induced activation of ERK in MG-63 cells. (A) MG-63 cells were stimulated with 100 µg/mL EMD for the indicated times at 37°C. Cells were harvested, and lysates were resolved in 10% SDS-PAGE and then transferred to a PVDF membrane. The membrane was immunoblotted with anti-phospho-p44/42 antibody (upper panel) and then stripped and immunoblotted with anti-p44/42 antibody (middle panel). Molecular markers (kDa) are shown in the left column. (B) MG-63 cells were treated for 30 min with U0126 (5 and 10 µM) before stimulation with 100 µg/mL Emdogain for 5 min. Cells were harvested, and lysates were resolved in 10% SDS-PAGE and then transferred to a PVDF membrane. The membrane was immunoblotted with anti-phospho-p44/42 antibody (upper panel), and then stripped and immunoblotted with anti-p44/42 antibody (middle panel). Molecular markers (kDa) are shown in the left column. Quantification of phosphorylation of p44/42 was performed densitometrically and corrected to the amount of total p44/42 protein. The intensities of each band are depicted as percent of maximum value (lower panel). These data are representative of more than 3 independent experiments.