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. 2004 Oct;24(20):9207–9220. doi: 10.1128/MCB.24.20.9207-9220.2004

FIG. 2.

FIG. 2.

Artemis is phosphorylated in vivo after exposure of MCF-7 cells to IR or UV irradiation. (A) Schematic of Artemis indicating locations of S/TQ motifs. Asterisks above or below the schematic indicate motifs conserved or not conserved in humans and mice, respectively. aa, amino acids. (B) Immunoblotting shows the absence of Artemis in Artemis-deficient P11 cells. The asterisk indicates a nonspecific loading control. (C) Treatment with either IR or UV results in phosphorylation of Artemis (Artemis-P). Artemis was immunoprecipitated from extracts of cells exposed to IR (10 Gy) or UV (20 J/m2) and subsequently incubated for 2 h (lanes 1 and 4). The same immunoprecipitate was incubated with alkaline phosphatase (lanes 2 and 5) or treated with alkaline phosphatase in the presence of the phosphatase inhibitor Na3VO4 (lanes 3 and 6). The immunoprecipitate from untreated cells (UT) is shown in lane 7. (D) Phosphorylation of Artemis as a function of IR dose examined 2 h after treatment. (E) Kinetic analysis of the phosphorylation of Artemis after the cells were exposed to IR (10 Gy) for times ranging from 5 min (5m) to 17 h. (F) Phosphorylation of Artemis as a function of UV dose examined 2 h after treatment. (G) Kinetic analysis of the phosphorylation of Artemis after exposure of cells to UV (20 J/m2).