Arf heterozygosity does not rescue the Mdm2 haploinsufficiency induced delay in tumor development.

<p>Kaplan-Meier survival curves of the indicated genotype of mice. The numbers of mice in each group are denoted by the <i>n</i> values.</p

Comparison of tumor spectrum in <i>Mdm2+/+</i> and <i>Mdm2+/−</i> mice with deficiencies in <i>Arf</i> and/or <i>p53.</i>

<p>Percentages were calculated from the total number of mice analyzed; – indicates no tumors of that type; F = female, M = male.</p

The contribution of p53 to the delay in tumorigenesis of <i>Arf−/−</i> mice caused by <i>Mdm2</i> heterozygosity.

<p>Kaplan-Meier survival curves of the indicated genotype of mice. The numbers of mice in each group are denoted by the <i>n</i> values. P values calculated by log-rank test.</p

The gene dosage of <i>Arf</i> contributes to tumorigenesis in the absence of <i>p53</i>.

<p>Kaplan-Meier survival curves of the indicated genotype of mice. The numbers of mice in each group are denoted by the <i>n</i> values. P values calculated by log-rank test.</p

Sarcoma subtype primarily dictated by Arf and p53 and not Mdm2.

<p>Percentage of total sarcomas from the specified genotype with the specific sarcoma subtype.</p

<b>Regression analysis of miRNA in non-recurrent and recurrent tumor relative to their normal adjacent tissue.</b>

<p>After normalizing miRNA expression in each adenocarcinoma to endogenous control RNU48, they were normalized to the expression in the corresponding matched adjacent normal control lung tissue. Limma <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101802#pone.0101802-Smyth1" target="_blank">[9]</a> was then used to identify differentially expressed miRNA between tumors that did and did not recur and calculate p values.</p

Validation of reduced miR-34b and miR-106b* levels in stage I lung adenocarcinoma and normal adjacent lung tissue.

<p>miR-34b and miR-106b* expression was measured using TaqMan qRT-PCR in a second cohort of stage I lung adenocarcinoma and adjacent normal lung tissue. ΔCt values graphed are relative to the endogenous small RNA control RNU48 with SEM. (A) Mean relative ΔCt value of tumors that did and did not relapse (n = 8) compared to mean value of adjacent normal lung tissues for both tumors (n = 7), *p = 0.03. (B) Mean relative ΔCt values of recurrent lung adenocarcinoma (n = 3) or adjacent normal lung tissue (n = 3), *p = 0.02. (C) Mean relative ΔCt values in tumors that did (n = 3) or did not (n = 5) relapse, *p = 0.02. (D) Mean relative ΔCt values of adjacent normal lung tissue from patients that did (n = 3) or did not relapse (n = 4), *p = 0.05. All p values calculated by student's t-test.</p

<b>Comparisons of miRNA expression between lung adenocarcinomas that did or did not recur and adjacent normal lung tissue.</b>

<p>miRNA expression was normalized to endogenous control RNU48 and fold change and p values of the differences were calculated by limma [log2(mean value group A/mean value group B)] <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101802#pone.0101802-Smyth1" target="_blank">[9]</a>.</p

<b>Clinical characteristics of patients with lung adenocarcinoma in this study.</b>

<p>*The year the tumor was resected from which the sample for analysis was obtained and frozen.</p

Reduced let-7b and -886-5p and increased miR-211 and -589 are potential biomarkers for stage I lung adenocarcinomas that recur.

<p>miRNA expression for let-7b, miR-886-5p, -211, and -589 was determined by TaqMan qPCR arrays in lung adenocarcinoma tumors that relapsed (n = 8) or that did not relapse (n = 6) and normal adjacent lung tissue for both (n = 8 and 6, respectively). ΔCt values graphed are relative to small RNA endogenous control RNU48. Lines connect patient matched samples.</p