The frequencies (percentages) of the five types of cancer- and normal tissue-specific alternative splicing.

<p>(a) 16 types of human cancer and 17 normal tissues, (b) the average values between tumors and normal tissues. The five colors indicate the five types of tissue-specific alternative splicing: cassette alternative exon, alternative 5′ splice site, alternative 3′ splice site, intron retention, and mutually exclusive alternative exons. Yellowish regions indicate over 30% of the frequencies.</p

A schematic representation of cancer-specific alternative gene splicing.

<p>(a) Brain cancer (gene ACYl), (b) breast cancer (SRP19), (c) liver cancer (CDK5), (d) lung cancer (CDKN1A), and (e) prostate cancer (SMS). Cancer-specific isoforms are showed on the bottom in each panel. The biological processes of these transcripts (GO process) are indicated on the right. Deleted domains are shown with blue arrows. Arrows with a right angle indicate the start codon, ATG.</p

Oncogenes and tumor suppressors with cancer-specific AS events.

<p>Oncogenes and tumor suppressors with cancer-specific AS events.</p

Numbers of cancer-specific AS transcripts and their genes.

<p>Numbers of cancer-specific AS transcripts and their genes.</p

Percentages of the types of alternative splice sites.

<p>The splice sites include GT-AG, GC-AG, GG-AG, GT-GG, and the others (a) in human cancer (b) and normal tissues. (c) Percentage distribution of the splice sites in five types of cancer and normal tissues (brain, breast, lung, liver, and prostate).</p

Immunohistochemical staining of RASSF1A in human testis cancer.

<p>Normal testis sample (<b>A</b>) and testis tumor with methylated RASSF1A (<b>B</b>). Normal testis section stained positively for RASSF1A. RASSF1A was mainly expressed in cytoplasm of the spermatogonia (SG), spermatocytes (SC), Sertoli cells (Sn) and Leydig cells (LC) of normal testis, while seminoma sample with methylated RASSF1A showed weak staining. Nuclei were stained with Hematoxlin.</p

Numbers of libraries and ESTs in normal tissues.

<p>Numbers of libraries and ESTs in normal tissues.</p

Numbers of normal tissue-specific AS transcripts and their genes.

<p>Numbers of normal tissue-specific AS transcripts and their genes.</p

Promoter activity and methylation analysis of <i>RASSF1A</i> in human testis cancer.

<p><b>A</b>, <i>RASSF1A</i> promoter activity by GFP expression under the control of different length of RASSF1A promoter segments. A predicted p53 binding site in <i>RASSF1A</i> promoter (−2718 bp) was showed in the construction c. <b>B</b>, CpG island prediction of <i>RASSF1A</i> by software MethPrimer online (<a href="http://www.urogene.org/methprimer/" target="_blank">http://www.urogene.org/methprimer/</a>). Predicted CpG island is indicated by blue color. <b>C</b>, Representative results of the methylation-sensitive PCR (MSP) analysis of <i>RASSF1A</i> in both seminoma and nonseminoma. M, MSP PCR; U, unmethylation-sensitive PCR. <b>D</b>, DNA methylation status of individual CpG sites by sodium bisulfite sequencing analysis. Black and white circles represent methylated and unmethylated CpGs respectively.</p

Down-regulation of <i>RASSF1A</i> by p53.

<p><b>A</b>, <i>RASSF1A</i> promoter activity was inhibited by p53 protein with a dose dependent manner. pRASSF1A-GFP (construction c in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0017017#pone-0017017-g001" target="_blank">Fig. 1</a>) was co-transfected with different concentration of CMV-p53 into COS-7 cells, and transfected cells were analyzed by the flow cytometry. <b>B</b>, pRASSF1A-GFP was transfected into COS-7 cells (<b>a</b>) or co-transfected with p53 (<b>b</b>), and images were taken under fluorescent microscopy. p53 remarkably decreased expression of RASSF1A-GFP. GFP was mainly expressed in the cytoplasm. Nuclei were stained with Hoechst. <b>C</b>, Overexpression of p53 in Siha cells decreased RASSF1A protein levels, revealed by Western blot analysis of RASSF1A and p53 expression using anti-RASSF1A or p53 antibody after transfection of the p53 expression plasmid into Siha cells. Lysates of non-transfected cells and cells transfected with the pcDNA3 vector were used as controls. β-actin was used as an internal control. Molecular weights of the proteins are shown on the right.</p