The indicated amounts (0.05 – 1 ug) of synthetic phosphorylated peptides (P) and their non-phosphorylated counterparts (N) were spotted onto nitrocellulose. The peptides were immunlotted with 1 ug/ml affinity purified antibody preincubated in the presence or absence of phosphorylated immunogen or non-phosphorylated peptides.

GFP-DBNL and the indicated single and double alanine mutants of these proteins were immunoprecipitated on GFP-Trap beads from transfected cells and then immunoblotted with anti-DBNL phospho Ser 269, anti-DBNL phospho Thr 291 and anti-GFP antibodies.