<i>RMRP</i> depletion is embryonic lethal.

<p><i>RMRP</i> depletion is embryonic lethal.</p

<i>RMRP</i> depletion leads to reduced levels of <i>RMRP</i> transcript.

<p>Total RNA was produced from E13.5 MEFs and <i>RMRP</i> level was measured by A. qRT-PCR B. Northern blot using either a sense or antisense <i>RMRP</i> probe. Error bars represent SD of three replicas.</p

Targeting of murine <i>RMRP</i>.

<p>A. Murine targeting vector (MTV) B. Southern blot of ES cells following selection for alleles with integrated MTV (the southern probe is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0026270#pone-0026270-g001" target="_blank">figure 1a</a>) C. PCR analysis of RC (<i>RMRP</i> conditional) pups D. PCR analysis of pups derived from the interbreeding of RC mice and mice expressing CMV-Cre.</p

<i>RMRP</i> depletion is lethal in early stage embryos.

<p><i>RMRP</i> depletion is lethal in early stage embryos.</p

Genes near <i>RMRP</i> are not essential for cellular viability.

<p>MEFs from E13.5 mice of either A. WT or B. <i>RMRP</i>+/− were transfected with siRNAs targeting <i>Ccdc107</i> or <i>E130</i>. Three days later RNA was extracted from the cells and qRT-PCR was preformed using primers for <i>RMRP</i>, <i>Ccdc107</i> or <i>E130</i>. C. The same cells as in A and B were plated (5000 cells/well) in a 96 well plate and 7 days post transfection cell number was assessed by Cell titer glow. Error bars represent SD of three replicas.</p

Analysis and Comparison of Somatic Mutations in Paired Primary and Recurrent Epithelial Ovarian Cancer Samples

<div><p>The <i>TP53</i> mutations have been proved to be predominated in ovarian cancer in a study from The Cancer Genome Atlas (TCGA). However, the molecular characteristics of recurrent ovarian cancers following initial treatment have been poorly estimated. This study was to investigate the pattern of somatic point mutations in matched paired samples of primary and recurrent epithelial ovarian cancers, using the OncoMap mutation detection protocol. We have adapted a high-throughput genotyping platform to determine the mutation status of a large panel of known cancer genes. OncoMap v.4.4 was used to evaluate genomic DNA isolated from a set of 92 formalin-fixed, paraffin-embedded (FFPE) tumors, consisting of matched paired samples of initially diagnosed and recurrent tumors from 46 epithelial ovarian cancer (EOC) patients. Mutations were observed in 33.7% of the samples, with 29.3% of these samples having a single mutation and the remaining 4.3% having two or more mutations. Among the 41 genes analyzed, 35 mutations were found in four genes, namely, <i>CDKN2A</i> (2.2%), <i>KRAS</i> (6.5%), <i>MLH1</i> (8.2%) and <i>TP53</i> (20.7%). <i>TP53</i> was the most frequently mutated gene, but there was no correlation between the presence of mutation in any gene and clinical prognosis. Furthermore, somatic mutations did not differ between primary and recurrent ovarian carcinomas. Every mutation present in recurrent samples was detected in the corresponding primary sample. In conclusion, these OncoMap data of Korean EOC samples provide that somatic mutations were found in <i>CDKN2A</i>, <i>KRAS</i>, <i>MLH1</i>, and <i>TP53</i>. No differences in mutational status between primary and recurrent samples were detected. To understand the biology of tumor recurrence in epithelial ovarian cancer, more studies are necessary, including epigenetic modifications or additional mutations in other genes.</p></div

Frequency of mutations.

<p>Frequency of mutations.</p

Clinicopathologic characteristics of patients.

†<p>Serous adenocarcinoma and transitional cell carcinoma in 3 cases, serous adenocarcinoma and mucinous carcinoma in 1 case, and transitional cell carcinoma and poorly differentiated carcinoma in 1 case.</p>‡<p>CR = complete remission; PR = partial response; SD = stable disease; PD = progressive disease.</p><p>*DFS = disease-free-survival.</p

Comparison of disease-free-survival and overall survival with respect to TP53 mutation.

<p>Disease-free-survival and overall survival are not different between <i>TP53</i> mutation-negative group and <i>TP53</i> mutation-positive group. A and B, Disease-free-survival and overall survival in total EOC patients (n = 46, 36 <i>TP53</i> mutation (−) vs. 10 <i>TP53</i> mutation (+)). C and D, Disease-free-survival and overall survival in serous adenocarcinoma (n = 35, 27 <i>TP53</i> mutation (−) serous vs. 8 <i>TP53</i> mutation (+) serous).</p

Concordance between primary and recurrent paired samples.

†<p>P means the tumor sample from primary tumor and R means the tumor sample from recurrent tumor.</p><p>Percentage showed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099451#pone-0099451-t004" target="_blank">table 4</a> means the ratio of mutant allele versus sum of wild and mutation for each position, that is to say the mutant allele frequency.</p