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. 2003 Dec;23(23):8601–8613. doi: 10.1128/MCB.23.23.8601-8613.2003

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FIG. 2. — WS cells are deficient in the poly(ADP-ribosyl)ation of cellular proteins after H₂O₂ treatment. (A) Immunofluorescence. Normal and WS primary fibroblasts (arrested in early S phase [B]) were incubated with H₂O₂ (500 μM) for 10 min at 37°C. Fixed cells were probed with anti-PAR antibodies. Control panels correspond to untreated cells. Nucleus staining by Hoechst 33342 is shown. (B) Synchronization in early S phase (see Materials and Methods) prior to H₂O₂ treatment is confirmed by propidium iodide staining and flow cytometry analysis. Arrows, G₁-S phase boundary. (C and D) Time course of PAR polymer formation. After H₂O₂ treatment, normal (C) and WS (D) primary cells were fixed at the indicated times and processed as for panel A. Similar results were obtained with two different WS primary cell lines (AG03141C and AG00780H) and two normal primary cell lines (MRC-5 and WI-38).

FIG. 2. — WS cells are deficient in the poly(ADP-ribosyl)ation of cellular proteins after H₂O₂ treatment. (A) Immunofluorescence. Normal and WS primary fibroblasts (arrested in early S phase [B]) were incubated with H₂O₂ (500 μM) for 10 min at 37°C. Fixed cells were probed with anti-PAR antibodies. Control panels correspond to untreated cells. Nucleus staining by Hoechst 33342 is shown. (B) Synchronization in early S phase (see Materials and Methods) prior to H₂O₂ treatment is confirmed by propidium iodide staining and flow cytometry analysis. Arrows, G₁-S phase boundary. (C and D) Time course of PAR polymer formation. After H₂O₂ treatment, normal (C) and WS (D) primary cells were fixed at the indicated times and processed as for panel A. Similar results were obtained with two different WS primary cell lines (AG03141C and AG00780H) and two normal primary cell lines (MRC-5 and WI-38).