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Fig. 4. UV-exposure induces EGFR mobility shift. (A) Cells were either left
untreated (lane 0) or exposed to 200 J/m2 UV (lanes 1,3) or 5 nM
EGF (lanes 2,4), and incubated for 1 hour (lanes 1,2) or 2 hours (lanes 3,4).
Cell lysates were subject to SDS-PAGE and western immunoblotting with
anti-EGFR. Unstimulated cells were used as controls. (B) Cells were either
left untreated, or exposed to 200 J/m2 UV or 5 nM EGF, and
incubated for 10 minutes before immunoprecipitation with anti-EGFR. The
immunoprecipitates were incubated in DEA buffer with or without alkaline
phosphatase (AP). The immunoprecipitates were subject to SDS-PAGE and western
immunoblotting with either anti-pY1173 or anti-EGFR. EGFR immunoprecipitates
from unstimulated cells were incubated in DEA buffer and used as controls. C,
control cells. The molecular weight markers presented at the left indicate 220
kDa.