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. 2004 Dec 10;6(1):70–76. doi: 10.1038/sj.embor.7400301

Figure 1.

Figure 1

Phosphorylation of SMN and pICln in vivo. (A) Extracts from metabolically labelled HeLa cells were immunoprecipitated with anti-pICln and anti-SMN antibody, separated by SDS–PAGE and analysed by autoradiography (lanes 1 and 2). The arrows indicate phosphorylated SMN and pICln, and the asterisk indicates an unknown phosphorylated product. Coomassie-stained proteins of isolated complexes are shown on the right. Components of the SMN and the pICln complex are indicated (# and +, respectively). (B) Phosphoamino-acid analysis of immunoprecipitated and metabolically labelled SMN and pICln. In-vivo-labelled proteins shown in (A) were excised from the membrane and hydrolysed. The sample was mixed with nonlabelled phosphoamino acids and separated by 2D-TLC. In-vivo-labelled amino acids were detected by autoradiography (panels b and d) and compared with the ninhydrin-stained standard (panels a and c). The asterisks denote the partially hydrolysed protein.