Relationship between ANA-staining intensity and histological grade of tumor (high, moderate, and poorly differentiated).

<p>While grading had no significant relationship with ANA-positivity of any specificity the presence of a speckled ANA pattern was significantly correlated with moderate-poor differentiation grade when compared with highly differentiated tumors (p = 0.04).</p

The frequency of different Antinuclear Antibodies (ANA)-patterns in sera from patients with epithelial ovarian cancer and benign pelvic conditions.

<p>Fisher's exact test was used to obtain <i>p</i>-values. Some sera contain ANAs of more than one specificity. Centr., centromere; Nuclear membr., nuclear membrane; Homog., homogeneous; cytopl., cytoplasmic; NS, non significant.</p

Reproducibility of mutant allele burden determination by the qPCR assays.

<p>Histogram plots showing reproducibility of percentage mutant allele burden in six separate qPCR runs. The mutant allele burden was determined for 11 different DNA samples from four different patients as indicated. The <i>JAK2</i> exon 12 mutations include K539L (PV6), V536-I546dup11 (PV4), and N542-E543del (PV1 and PV2) in high, intermediate and low levels of mutant allele burden. The data is presented as percentage mean values ± standard deviation (SD).</p

No correlation between serum-CA-125 values and the presence and intensity of ANA.

<p>The immunofluorescence score is depicted as a function of CA-125 values for the samples from benign (red symbols) and malignant cases (green symbols). Cut-off for positivity in the CA-125 test is 35 U/mL. All patients with benign ovarian tumors and epithelial ovarian cancers are included in this figure.</p

Antinuclear Antibodies (ANA) specificities.

<p>Examples of ANA-patterns of strongly positive sera staining in a cytoplasmic (A) and a speckled (B) pattern. In (C) is shown the distribution of the main types of ANA patterns as well as the signal strength in the benign and the malignant group. Strongly positive samples are almost only seen in the group with malignancy.</p

Amplification plots of dilution series and standard curve of qPCR assay.

<p>A: Representative amplification plots of plasmid containing <i>JAK2</i> exon 12 mutation diluted into wildtype genomic DNA detected by the mutation specific assay. The ten fold dilution series starting at 15,000 copies is continued as two-fold dilutions after 15 copies down to 1.9 copies as indicated. B: Representative standard curve for the mutation and wildtype assays producing correlation coefficients >0.990. The average slope of the standard curves was −3.4.</p

Study demographics in patients diagnosed with borderline ovarian tumor, ovarian cancer or a benign ovarian tumor.

<p>*endometrioid adenocarcinoma N = 11, Clear cell neoplasms N = 6 and carcinosarcoma N = 4.</p><p>**No significant difference for the subset of 127 matched patients with benign conditions: Median age: 64 (range 54–90).</p

Primer and probe sequences for <i>JAK2</i> exon 12.

<p>Primers used for qPCR screening and quantitative determination consisting of a common forward primer and probe in addition to a mutation specific reverse primer. The primers listed for quantitative determination were only used exclusively for determination of mutant allele burden. Lowercase letters in sequences indicate intended mismatches.</p

Identification of <i>JAK2</i> exon 12 mutations.

<p>A: Sequence of F537-I540delinsLV mutation of PV4 revealing a 10 base-pair deletion and a four base-pair insertion. B: Difference plot of high resolution melting (HRM) analysis detecting N542-E543del, F537-I540delinsLV and K539L <i>JAK2</i> exon 12 mutations in PV1, PV3, PV4, PV5 and PV6. C: Phase-contrast microphotograph showing Epo-independent growth of endogenous erythroid colonies (EEC) at day 14 of culture. Scalebar: 200 µm. HRM, high resolution melting.</p

Mutant <i>JAK2</i> exon 12 allele burden in bone marrow and peripheral blood cell lineages.

<p>Histogram plot displaying the <i>JAK2</i> exon 12 mutation burden in bone marrow, peripheral blood, CD16⁺ granulocytes, CD14⁺ monocytes, CD3⁺ T-lymphocytes, and CD19⁺ B-lymphocytes in patients PV1-PV6. PV2 had very low level of <i>JAK2</i> exon 12 mutant allele burden except for the bone marrow sample. PV1-PV3 and PV5 appeared to be heterozygous, whereas PV4 and PV6 were homozygous. Note that DNA from bone marrow samples could not be obtained from patients PV5 and PV6. BM, bone marrow and PB, peripheral blood.</p