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. 2010 Jan 15;38(9):2891–2903. doi: 10.1093/nar/gkp1250

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — ITPA-deficient primary MEFs exhibit various cellular dysfunctions. (A) ITPA deficiency caused a significantly increased accumulation of inosine in cellular RNA. Inosine level was determined by LC–MS/MS analysis of cellular RNA prepared from embryos (N3). Result of non-repeated measures ANOVA (two-tailed), P = 1.69 × 10⁻⁷. Student–Newman–Keuls (SNK) post hoc test, **P < 0.01 (versus Itpa^+/+ and Itpa^+/−). Data are shown as the mean ± SD (n = 3 independent embryos). (B) ITPA deficiency caused a significantly increased accumulation of deoxyinosine (dI) in nuclear DNA. Deoxyinosine (dI) level was determined by LC–MS/MS analysis of nuclear DNA prepared from embryos (N3). Result of non-repeated measures ANOVA (two-tailed), P = 0.00038. SNK post hoc test, **P < 0.01 (versus Itpa^+/+ and Itpa^+/−). Data are shown as the mean ± SD (n = 3 independent embryos). (C) ITPA deficiency impairs normal cell proliferation. Primary MEFs (Passage 2) isolated from four separate Itpa^−/– embryos showed significant prolonged doubling time in comparison to those from Itpa^+/+ and Itpa^+/− embryos. Result of repeated measures ANOVA (two-tailed), P = 0.0005. Bonferroni/Dunn post-hoc test, *P < 0.05 (versus Itpa^+/−), **P < 0.01 (versus Itpa^+/+). Data are shown as the mean ± SD (n = 4 independent MEFs). (D) ITPA deficiency causes G2/M arrest. Primary MEFs (Passage 5) were subjected to flow cytometry analysis and the percentages of Cell-cycle phases in each MEF set were determined. Result of non-repeated measures ANOVA (two-tailed), P = 1.74 × 10⁻⁸. Bonferroni post hoc test, **P < 0.01 (versus Itpa^+/+ and Itpa^+/−). Data are shown as the mean ± SD (n = 3 independent isolates).