MtDNA content in the lung cancer patients.

<p>mtDNA content was measured by multiplex real-time PCR using nuclear encoded β-actin and mitochondria encoded COI gene. A ratio of COI/β -actin corresponds to the fold difference compared to control. MtDNA content increased significantly (P<0.001) in the mucosal biopsies and corresponding tumors compared to the normal control in both the patients as indicated. N: Normal lymph node; M: Mucosa; T: Tumor. *P value <0.05 compared to normal lymph node.</p

Pattern of mtDNA mutation in patient 2.

<p>The patient was operated on 2005 for surgical removal of tumor from the right upper lobe. Five bronchoscopically abnormal airway mucosal biopsies were obtained from main carina (M1), right lower lobe (M2 and M3) and right upper lobe (M3 and M4) surrounding the tumors as depicted (Panel A–E). The mucosal biopsies exhibited a range of 2–5 mtDNA mutations as represented in different color (Panel A–E). Identical mutations are shown in same color in different biopsies and the tumor. A different color code was also used for each mucosa to indicate the clonal progression of the lesions with accumulated mtDNA mutations. Representative histological photomicrograph of the tumor and normal mucosa are shown in Panel A and D. The tumor exhibited 9 mtDNA mutations (Represented in the Bottom rectangle) including all the 8 mutations exhibited by the five mucosal biopsies (Matched color). An additional mtDNA mutation (<i>T7001C</i>) detected in the tumor is underlined in black in the bottom rectangle. Tumor was encircled to indicate its development from the clonal mucosal patches. M: Mucosa.</p

Pattern of mtDNA mutation in patient 3.

1<p>Revised Cambridge Reference Sequence;</p>2<p>Mucosal biopsies from 5 different regions were pooled together because of small sample size;</p>3<p>Tumor adjacent histologically normal margin.</p

Pattern of somatic mtDNA mutation in the 8 lung cancer patients.

1<p>RCRS: Revised Cambridge Reference Sequence;</p>2<p>Normal Lymphocytes.</p

The pattern of germ line mtDNA sequence variants in the lung cancer patients.

1<p>Revised Cambridge Reference Sequence;</p>2<p>Matched normal lymphocytes;</p>3<p>R: Reported; N: Novel as per Human Mitochondrial Database and relevant literature.</p

MtDNA genetic mutation tree in patient 1.

<p>Possible clonal evolution of the lung tumors has been depicted. Each mucosal biopsy (Circled, M1-M5) shared some identical mtDNA mutations (Matched color) along with additional mutations in the heterogeneous mucosal field. The fittest clones emerged in the primary tumors (T1-T2) with the more selective mtDNA mutations (Total 20 mtDNA mutations, 16 were shared between M1-M5 as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006533#pone-0006533-g001" target="_blank">Figure 1</a>).</p

History of the 8 lung cancer patients additionally sequenced for mtDNA mutation.

1<p>RUL: Right upper lobe; RLL: Right lower lobe; LLL: Left lower lobe;</p><p>LUL: Left lower lobe; RML; Right middle lobe.</p>2<p>SCC: Squamous Cell Carcinoma;</p>3<p>All the tobacco positive patients have a smoking history of more than 10 years;</p><p>NS: Non Smoker; PYH: Pack year smoked.</p

mtDNA content index in patient 3.

<p>mtDNA content increased significantly (P<0.05) in the mucosa (M), adjacent normal (NT) and in the metaplastic margin (MT) compared to normal lymph node used a s control.</p

Frequency of positive cases [n (%)] and distribution of Methylation Quotient levels [Mean (range)] in normal parotid tissue, PA, ACC, MEC, and SDC.

<p>*Note: Frequency of positive cases is expressed as number and (%). Distribution of MQ levels is the ratio of the methylation of the gene to β-actin ×1,000.</p

Methylation by tissue and tumor type.

<p>Overview of the log methylation quotients of normal (N), benign pleomorphic adenoma (PA), and cancerous (CA) salivary gland tumors from the 19 tumor suppressor genes tested.</p