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

Phylogenetic tree, amino acid alignments, domains and modification sites of Spata33.

<p>A). Phylogenetic tree of Spata33 in mammals. Phylogenetic analysis was performed with Phylip. Numbers on the branches represent the bootstrap values from 1000 replicates obtained using the Neighbor-Joining method. The scale bar corresponds to the estimated evolutionary distance units. GenBank accession numbers are as follows: <i>Callithrix jacchus</i>, XP_002761326.1; <i>Canis lupus familiaris</i>, XP_003434741.1; <i>Cricetulus griseus</i>, XP_003495126.1; <i>Gorilla gorilla gorilla</i>, XP_004058209.1; <i>Homo sapiens,</i> BAG64150.1; <i>Macaca mulatta,</i> XP_001104069.2; <i>Mus musculus</i>, NP_796253.2; <i>Nomascus leucogenys</i>, XP_003280677.1; <i>Otolemur garnettii,</i> XP_003800884.1; <i>Pan paniscus</i>, XP_003805830.1; <i>Pan troglodytes</i>, XP_511172.4; <i>Papio anubis,</i> P_003917381.1; <i>Rattus norvegicus</i>, NP_001099665.1; <i>Saimiri boliviensis boliviensis</i>, XP_003944616.1; <i>Tupaia_chinensis</i>, ELW62868.1. B). Alignment of amino acid sequences of the Spata33 proteins. Amino acids that are identical in all these species are shown in white letters on black background. The DUF4609 domain predicted by Pfam is boxed. C). Schematic mapping of potential protein domains and post-translational modification sties. The predicted sites for N-myristoylation, N-glycosylation and phosphorylations in Spata33 were indicated (N-Myr, N-myristoylation site; N-Glyc, N-glycosylation site; S/T, Serine/Threonine phosphorylation sites).</p

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

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 cancer-specific AS transcripts and their genes.

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

Oncogenes and tumor suppressors with cancer-specific AS events.

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

Immunostaining of Spata33 protein in the juvenile (aged 10–20 days) and mature testes (aged 35 days) using anti-Spata33 antibody (A), and subcellular localization of Spata33 protein in the GC-1 cells and TM4 cells (B).

<p>A). Spata33 was expressed mainly in the spermatogonia (white arrow heads), spermatocytes (red arrows) and round spermatids (red arrow heads). The signals were observed in both the cytosol and nuclei from P12-P35, whereas no signals were detected in P10. In controls, no positive signals were observed in P35 testis sections when Spata33 antibody was replaced by 1% normal rabbit serum. Nuclei were re-dyed with Hematoxylin (blue). Bars, 20 µm. B). GC-1 and TM4 cells were transfected with Spata33-GFP (green, Excitation 488 nm, Emission 507 nm) and stained with Hoechst (blue, Excitation 352 nm, Emission 461 nm) respectively. Spata33 is localized both in the nuclei and cytoplasm within GC-1 cells or TM4 cells. Bar, 10 µm.</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

Numbers of libraries and ESTs in normal tissues.

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