Data from: Increased sensitivity of DNA damage response-deficient cells to stimulated microgravity-induced DNA lesions

Li N, An L, Hang H

Date Published: April 29, 2015

DOI: http://dx.doi.org/10.5061/dryad.3vc83

 

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Title Fig1-A Neutral comet assay MES cell
Downloaded 9 times
Description Fig. 1. Effects of SMG on DNA damage and apoptosis in Rad9+/+ and Rad9-/- MES cells (A) Evaluation of DNA double strand break by neutral comet assay in Rad9+/+ and Rad9-/- MES cells cultured under 1G or SMG condition. Time points were 1, 2, 3, 4 and 5 days. At least 50 cells for each datum were scored for comet tail moment.
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Title Fig1-B
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Description Flow cytometric analysis of γ-H2AX formation in Rad9+/+ and Rad9-/- mMES cells cultured under 1G or SMG condition for 1 or 5 days
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Title Fig1-C
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Description Evaluation of DNA damage by alkaline comet assay in Rad9+/+ and Rad9-/- MES cells cultured under 1G or SMG condition for 1 or 5 days. At least 50 cells of each datum were scored for comet tail moment.
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Title Fig1-D
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Description Evaluation of DNA damage by alkaline comet assay in Rad9-/- MES cells with ectopic expression of Rad9 (Rad9-/-+Rad9 MES cells) cultured under 1G or SMG condition for 1 or 5 days. At least 50 cells of each datum were scored for comet tail moment.
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Title Fig1-E
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Description Flow cytometric analysis of Rad9+/+ and Rad9-/- MES cells cultured under 1G or SMG condition for 1 day to assess apoptosis using Annexin V labeling. Experiments were performed thrice and representative analyzes are shown (upper). The lower part is the quantitative comparison of apoptosis between the 1G Group and the SMG Group.
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Title Fig2
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Description DNA double strand break by neutral comet assay in Mdc1+/+ and Mdc1-/- MEF cells cultured under 1G or SMG condition for 1 or 5 days. At least 50 cells of each datum were scored for comet tail moment.
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Title Fig3 a,b,c
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Description (A) Flow cytometric analysis of ROS activity in Rad9+/+ and Rad9-/- MES cells exposed to 1G or SMG condition for 1 or 5days. (B) Flow cytometric analysis of ROS activity in Rad9-/-+Rad9 MES cells exposed to 1G or SMG condition for 1 or 5days. (C) N-acetylcysteine inhibited SMG-induced increase of ROS formation in Rad9-/- MES cells. Rad9-/- MES cells were mock-treated or treated with 0.05, 0.1 or 0.5 mM N-acetylcysteine under SMG for 1 day.
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Title Fig3 d e
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Description (D) Evaluation of DNA damage by alkaline comet assay in Rad9-/- MES cells mock-treated or treated with 0.5 mM N-acetylcysteine under 1G of SMG condition for 1 day. (E) Evaluation of DNA damage by neutral comet assay in Rad9-/- MES cells mock-treated or treated with 0.5 mM N-acetylcysteine under 1G of SMG condition for 1 day.
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Title Fig4
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Description (A) Flow cytometric analysis of ROS activity in Mdc1+/+ and Mdc1-/- MEF cells exposed to 1G or SMG condition for 1 or 5days.(B) N-acetylcysteine inhibited SMG-induced increase of ROS formation in Mdc1-/- MEF cells. Mdc1-/- MEF cells were mock-treated or treated with 0.05, 0.1 or 0.5 mM N-acetylcysteine under SMG for 1 day. (C) Evaluation of DNA damage by neutral comet assay in Mdc1-/- MEF cells mock-treated or treated with 0.5 mM N-acetylcysteine under 1G of SMG condition for 1 day.
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Title Fig5
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Description (A) Quantitative real-time PCR analysis of Nox2 mRNA expression in Rad9+/+ and Rad9-/- MES cells exposed to 1G or SMG condition for 1 or 5days. The expression levels of Nox2 were normalized to the endogenous control GAPDH expression. (B) Quantitative real-time PCR analysis of Nox4 mRNA expression in Rad9+/+ and Rad9-/- MES cells exposed to 1G or SMG condition for 1 or 5days. The expression levels of Nox4 were normalized to the endogenous control GAPDH expression. (D) Quantitative comparison of Nox2 expression. Data were derived from three independent experiments . The expression levels of Nox2 protein were normalized to the endogenous control GAPDH protein expression.
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Title Fig6-A
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Description Histograms of superoxide dismutase enzyme activity
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Title Fig6-B
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Description Histograms of catalase enzyme activity
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Title Fig6-C
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Description Histogram of Glutahione peroxidase
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Title Fig7
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Description Quantitative comparison of Nox2 expression. Data were derived from three independent experiments The expression levels of Nox2 were normalized to the endogenous control GAPDH expression
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Title Fig8A
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Description (A) The initial MES cells seeding number was 3×104. The doubling generation curve was generated by dividing the cell number with 104 and then transferring the quotient to the logarithm to the base2.
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Title Fig8B
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Description The initial MEF cells seeding number was 105. The doubling generation curve was generated by dividing the cell number with 104 and then transferring the quotient to the logarithm to the base2.
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When using this data, please cite the original publication:

Li N, An L, Hang H (2015) Increased sensitivity of DNA damage response-deficient cells to stimulated microgravity-induced DNA lesions. PLOS ONE 10(4): e0125236. http://dx.doi.org/10.1371/journal.pone.0125236

Additionally, please cite the Dryad data package:

Li N, An L, Hang H (2015) Data from: Increased sensitivity of DNA damage response-deficient cells to stimulated microgravity-induced DNA lesions. Dryad Digital Repository. http://dx.doi.org/10.5061/dryad.3vc83
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