(A) Schematic representation of Pellino proteins indicating the location of the RING domain and of the 24 amino acid insert in Pellino3a (grey). Lines indicate the deletion variants. The Pellino amino acids in these GST-fusion proteins are indicated.
(B) The indicated GST-fusion of Pellino proteins or of deletion variants were incubated with HEK293T lysates that contained overexpressed IRAK1. Following washing, bound proteins were eluted by denaturation, analyzed by SDS-PAGE, and visualized by immunoblotting with α-Xpress antibody to detect IRAK1 and Coomassie blue staining to detect the GST-fusion proteins. “In” indicates 0.8 % of lysate. Positions of molecular mass markers are shown.