Figure 1.

Atg13 interacts with ULK1, ULK2, and FIP200. (a) ULK1 and ULK2 are coimmunoprecipitated with Atg13. HA-tagged ULK1 was expressed with myc-tagged Atg13 or control proteins (S6K1 and tubulin) in 293T cells. Anti-myc immune complexes were assessed for the presence of HA-ULK1 and HA-ULK2 by Western blotting. (b) Atg13 binds ULK1 and ULK2. Myc-tagged Atg13 was expressed with HA-tagged ULK1, ULK2, or control proteins (S6K1, tubulin) in 293T cells. Anti-HA immune complexes were isolated and the amount of Atg13 was assessed by Western blotting. (c) Recombinant Atg13 is coimmunoprecipitated with ULK1. Myc-tagged Atg13 or control proteins was expressed in 293T cells. Anti-myc immune complexes were isolated, and the presence of endogenous ULK1 was assessed by Western blotting. (d) Endogenous Atg13 is coimmunoprecipitated with ULK1. ULK1 immunoprecipitate was obtained from 293T cells in the presence (+pep) or absence (−pep) of ULK1 antibody epitope peptides and the amounts of Atg13 in the immune complexes were assayed by Western blotting. (e) Atg13 interacts with FIP200. Myc-tagged Atg13 or tubulin (control) was coexpressed with HA-FIP200 in 293T cells. Anti-myc immune complexes were analyzed for the presence of FIP200 on Western blots. (f) FIP200 interacts with Atg13, ULK1, and ULK2. HA-tagged Atg13, ULK1, and ULK2 were coexpressed with myc-tagged FIP200 in 293T cells. HA immunoprecipitates were analyzed for the amount of myc-FIP200 on Western blots.