Western blot analysis of MICOL24 cell line. (A) Representative Western blot analyses performed on the MICOL24 colon cancer cell line transfected either with the empty vector (mock) or pcDNA3.1-HsNEU3. Cells were treated for 3 h with 1 μg/mL cetuximab. Protein extracts were separated on a 12% SDS-PAGE and probed with anti-P-EGFR, anti-EGFR, anti P-Akt, anti-Akt, anti-P-ERK, anti-ERK and anti-PTEN antibodies. GAPDH and Vinculin were used as loading controls. The experiments were performed in triplicate. (B,C) Densitometric analysis was performed with Scion Image Software. Values are expressed by comparing the data obtained after cetuximab treatment with those obtained without cetuximab administration (B) by comparing the data obtained after transfection with NEU3 with those obtained after transfection with the empty vector (mock) (C). Values are presented as means ± standard error (SE). * p < 0.05, ** p < 0.01, *** p < 0.001.