FIGURE 2

Effect of gefitinib and cetuximab on primary human keratinocytes. Representative immunoblots of lysates from NHEK (A, C) and 22Rv1 cells (B) are shown. (A) NHEK and (B) 22Rv1 cells were serum-starved for 24 h and treated with increasing levels of gefitinib 2 h before stimulation with 4 nM EGF for 10 min at 37°C. (C) NHEK were serum-starved for 24 hrs in the absence or presence of 5 g ml^-1 cetuximab before stimulation with 4 nM EGF for 10 min at 37°C. Following stimulation cells were lysed in RIPA buffer and equal amounts of total protein were subjected to SDS-PAGE and transferred to PDVF-membranes. Blots were analyzed with specific antibodies to pEGFR, EGFR, pMAPK as indicated and reprobed for -actin to account for loading differences.