Figure 1.

Artemis interacts with DDB1 and DDB2 in vivo. (A) Gel stained with Coomassie blue showing proteins from a GST-Artemis pull-down experiment. GST-Artemis expressed in HEK293 cells was pulled-down by glutathione sepharose beads (left lane). A pull-down from untransfected cells was used as a control (middle lane). Labeled band indicate proteins identified by mass spectrometric analysis. “M” indicates marker proteins. (B) Immunoblot showing that Artemis co-IPs with DDB1 in the primary fetal lung cell line MRC5. (C) Immunoblot showing that endogenous DDB1 interacts with Artemis in vivo before and after UV or IR irradiation. HEK293 cells were treated with UV (25 J/m²), IR (10 Gy) or mock treated (UT). Cells were harvested for co-IP analysis two hours post-irradiation. “Pre” indicates pre-immune serum. (D) Reciprocal co-IP experiment showing that endogenous Artemis interacts with DDB1 in vivo before and after UV damage. HEK293 cells were treated with UV (25 J/m²) or left untreated. Cells were incubated for two hours after treatment and subsequently harvested for analysis. (E) Immunoblot showing that endogenous Artemis interacts with ectopically expressed DDB2. HEK293 cells were transfected with GST-DDB2, and 48 h later, cells were harvested for co-IP experiments. (F) Co-IP experiment showing the interaction between Artemis and DDB2. GsT-DDB2 and GFP-Artemis were co-transfected into HEK293 cells, and 48 h later, cells were harvested for analysis.