Figure 7.

Erk 1/2 phosphorylation in GN11 cells stimulated with anti- PrPc antibody. GN11 cells were transfected with empty HA-plasmid Prk7HA (lane 1) or PrP-HA plasmid (lanes 2–7) and incubated with anti-PrP-antibody (3F4) for 20 (lane 3) and 40 (lanes 4 and 7) min. In lane 5, cells were incubated with an unspecific antibody (anti-β-actin) for 40 min. In lanes 6 and 7, cells were treated with the Fyn kinase inhibitor PP2 (30 μm, 4 h). Cell lysates were collected and analyzed by western blot using anti-phospho-p44/42 MAP kinase antibody (a) and anti-β-actin antibody (b). Band intensities were evaluated by ImageMaster software (Pharmacia Biotech, Sweden). In (c), we reported the intensity values of the bands corresponding to phosphorylated Erk 1 and Erk 2 (black and white bars, resp.) normalized with respect to β-actin band intensity values. Data are representative of three independent experiments.