U2OS cells were split into two aliquots and cultured in one of
the following media (DMEM + 10 % dialysed FCS):
i) light medium: contains Arg-0 and Lys-0
ii) heavy medium: contains Arg-10 and Lys-8
Cells were passaged at least three times in the respective medium to ensure complete
labelling of proteins. SILAC experiments were performed in a forward and a reverse
reaction: for the forward experiment, heavy-labelled cells were irradiated with 20 Gy and
harvested 45 minutes after irradiation, while light-labelled cells were left untreated. In the
reverse experiment, labels were swapped and light-labelled cells were irradiated, while
heavy labelled cells were left untreated.