Figure 3

Mapping the P-STM phosphoepitope on lamin A/C using synthetic phosphopeptides. (A) Sequences and properties of the synthetic peptides used in this study. (B) GST-lamin C-N1 protein (0.5–2 μg), phosphorylated without or with CDK1, and synthetic peptides (10 μg) were subjected to dot-blot analysis with the P-STM antibody. (C) Lamins A and C were immunoprecipitated from total extracts (1 mg) of Noc-treated HeLa S3 cells and resolved by SDS-PAGE on 10% gels. Upper panel: In competition assays, the IP products were analyzed by immunoblotting with P-STM antibody in the presence of the indicated peptide (50 μg/mL). Lower panel: Immunoblot analysis of the same IP products using an anti-lamin A/C antibody (loading control).