Analysis Tools
cellular
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• retroviral cre-induced mutant MEFs show greater growth inhibition in response to gamma-irradiation (1-4 Gy) compared to control cells
• after irradiation, knockdown MEFs show a significantly higher level of sister chromatid unions (SCU) than control cells; other forms of chromosome damage are not significantly different from controls; at 8 hours, significantly more chromosomal aberrations (SCU) remain in mutant MEFs indicating impaired repair processes
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• cultured MEFs infected with a retroviral cre increase in number more slowly and display more DNA damage markers than MEFs transfected with a retroviral reporter
• after treatment with alphidicolin, cells display lower incorporation of BrdU than control MEFs
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• 4 and 8 hours after irradiation, double strand break (DSB) repair, assessed by presence of gammaH2AX foci, is impaired in Bccip knockdown (cre treated) MEFs compared with control cells (many more foci are observed indicating inefficient repair of breaks)
• numbers of Rad51 foci are reduced after 2 Gy of irradiation compared to number detected in irradiated wild-type cells
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• Bccip MEFs infected with a retroviral cre vector show significantly higher levels of spontaneous chromosome breakage (single chromatid breaks with unpaired fragments, paired sister chromatid fragments (pSCF) and sister chromatid unions (SCU)) than control MEF cultures; level of sister chromatid exchanges (SCE) is only modestly elevated
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homeostasis/metabolism
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• 4 and 8 hours after irradiation, double strand break (DSB) repair, assessed by presence of gammaH2AX foci, is impaired in Bccip knockdown (cre treated) MEFs compared with control cells (many more foci are observed indicating inefficient repair of breaks)
• numbers of Rad51 foci are reduced after 2 Gy of irradiation compared to number detected in irradiated wild-type cells
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