Figure 4.

ULK2 promotes degradation of BCL10. (a) HEK293T cells were transiently transfected with expression vectors encoding for the indicated polypeptides. About 24 h later, cell lysates were prepared and monitored for endogenous BCL10 expression by immunoblot assay. (b) HEK293T cells were transiently transfected with expression vectors encoding for the indicated polypeptides. About 24 h after transfection, cells were left untreated or treated with the lysosome inhibitor leupeptin (10 μM) for 1 h. Cell lysates were then prepared and monitored for conversion of LC3-I to LC3-II by immunoblot assay. (c) NHEK were transfected with an expression vector encoding for wt ULK2 or the kinase-dead mutant ULK2K39I. About 16 h later, cell lysates were prepared and BCL10 expression was determined by immunoblot assay. (d) Immunofluorescence analysis of BCL10 in NHEK infected with lentiviral vectors expressing the indicated polypeptides and left in medium alone or medium supplemented with the autophagy inhibitor 3-MA (5 mM). Data shown are representative of three independent experiments