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Fig. 8. (A) No mobility shift is observed for Dmnk/DChk2 in response to HU or IR, irrespective of the presence of Grp/DChk1. Western-blot analysis of Dmnk/DChk2 protein in S2 cells and Grp/DChk1-depleted S2 cells treated with 10 mM HU for 15 hours or treated with 150 Gy IR followed by 2 hours of recovery. In S2 cells, Dmnk/DChk2 protein was detected (lane 1) and, in response to HU (lane 2) or IR (lane 3), no shift in mobility of Dmnk/DChk2 was observed. (lane 4) Downregulation of Dmnk/DChk2 protein levels using RNAi. Depletion of Grp/DChk1 using RNAi did not affect the mobility of Dmnk/DChk2 protein in untreated (lanes 5,7), HU-treated (lane 6) or IR-treated (lane 8) S2 cells. As a loading control, {gamma}-tubulin protein levels were detected. (B) Phosphorylation of Grp/DChk1 in response to HU or IR is unaffected in Dmnk/DChk2-depleted cells. Western-blot analysis of Grp/DChk1 in S2 cells and Dmnk/DChk2-depleted cells treated with HU (10 mM, 15 hours) or IR (150 Gy, 2 hours recovery). In S2 cells and in Dmnk/DChk2-depleted cells, a slower-migrating form of Grp/DChk1 was detected in response to HU (lanes 2,6) or IR (lanes 4,8). As a loading control, {gamma}-tubulin protein levels were detected.





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