Radiation induced caveolin-1 linked EGFR internalization is triggered by src kinase. Confluent A549 cells were irradiated with 4 Gy (A). At the time points given, src protein was immune-precipitated from cytoplasmic protein fraction. Proteins were separated by SDS-PAGE and after blotting the protein amounts of src, caveolin1 and EGFR were quantified. The same procedure was applied after cell stimulation with EGF (B). Phosphorylation of EGFR at the tyrosine No. 845, 992 and 1173 was quantified with help of specific antibodies following a standard western procedure. Equal protein amounts for immune-precipitation were documented by showing actin expression within aliquots of input proteins. (C). Protein expression was quantified by densitometry and shown as fold-induction relative to untreated control. The mean protein expression derived from three experiments was given below each band.
Dittmann et al. Molecular Cancer 2008 7:69 doi:10.1186/1476-4598-7-69