Cdc5L is required for the activation of ATR‐mediated checkpoint pathways. Cdc5L is required for the activation of ATR‐mediated checkpoint pathways. (A,B)

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Cdc5L is required for the activation of ATR‐mediated checkpoint pathways. Cdc5L is required for the activation of ATR‐mediated checkpoint pathways. (A,B) Immunoblots showing that Cdc5L is required for the phosphorylation of Chk1 and Rad17 in HCT‐116 cells. The ultraviolet (UV) radiation dose and HU concentration were 20 J/m2 and 2 μM, respectively. Cells were fixed for analysis 1 h after ultraviolet radiation treatment and 3 h after the introduction of HU. (C–E) Knockdown of Cdc5L by siRNA in HeLa cells results in defective phosphorylation of Chk1, and monoubiquitylation of FancD2. In (E) ‘L’ and ‘S’ refer to the ubiquitinated and unmodified forms of FancD2, respectively. (F) Depletion of Prp19/Pso4 destabilizes Cdc5L and causes impairment of ATR activation. Experiments were carried out as described in (C); Ku70 is shown as a loading control. ATR, ataxia‐telangiectasia and Rad3‐related protein; Cdc5L, cell division cycle 5‐like protein; Chk1, checkpoint kinase 1; Ctrl, control; Dox, doxycycline; FancD2, Fanconi anaemia complementation group D2 protein; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; HU, hydroxyurea; MMC, mitomycin C; Prp19, precursor messenger RNA‐processing factor 19; Rad17, cell cycle checkpoint protein Rad 17; siRNA, small interfering RNA. Nianxiang Zhang et al. EMBO Rep. 2009;10:1029-1035 © as stated in the article, figure or figure legend