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Fig. 1. STAT-1 functions in the S-phase and G2/M checkpoint. (A) U3A cells lacking STAT-1 display an RDS phenotype. The method involves pre-pulsing 2fTGH, U3A or U3A-ST1 cells with ¹⁴C thymidine, irradiation, and then assessing ³H uptake after 2-5 Gy ${gamma}$ -irradiation (IR); DNA synthesis was assessed 2 hours later. The upper panel shows expression levels of STAT-1 (ST1) and also induction of phospho-STAT-1^Y701 (pST1) in response to ${gamma}$ -interferon for 30 minutes in 2fTGH, U3A or U3A-ST1 cells. (B) Dose effect of ionising ${gamma}$ -irradiation (1-10 Gy) and DNA synthesis in 2fTGH, U3A or U3A-ST1 cells. (C) Analysis of the G2/M checkpoint in 2fTGH, U3A or U3A-ST1 cells exposed to 10 Gy ${gamma}$ -irradiation (IR). The mitotic index of cells was assessed by histone H3 phosphorylation 4 hours after irradiation. (D) Analysis of the G2/M checkpoint in 2fTGH, U3A or U3A-ST1 cells exposed to 10 Gy ${gamma}$ -irradiation (IR), where the mitotic index of cells was assessed by DAPI staining of chromosomal metaphase spreads of treated versus untreated cells. (E) Cell survival was assessed following exposure to 10 Gy ${gamma}$ -irradiation in 2fTGH, U3A or U3A-ST1 cells for 72 hours. After Crystal Violet staining, the percentage of cell survival was determined. Data are representative of three separate experiments.

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