Figure S11. Western blot validation of chain length–dependent Ub interactors.

(A, B) Silver stain of Ub bait input after streptavidin immobilization for experiment in (B). * indicates streptavidin band from denaturation of streptavidin beads during sample prep for SDS–PAGE. (B) Western blot of Ub pulldown of MS-identified chain length–dependent interactors from the chloroacetamide- (top) and N-ethylmaleimide–treated (bottom) lysate. Pulldown blotted using anti-FAF1, anti-DDI2, anti-APPL1, anti-CCDC50, and anti-Ub antibodies. Input bait is Ub input before streptavidin immobilization. The same experiment as in Fig 5B.