Journal: PLoS ONE

Article Title: Evolutionary Diversification of SPANX-N Sperm Protein Gene Structure and Expression

doi: 10.1371/journal.pone.0000359

Figure Lengend Snippet: (a) RT-PCR analysis of the SPANX-N gene subfamily in normal adults tissues and cancer cell lines. cDNA was prepared from a panel of human tissue mRNAs and cell lines. Oligonucleotides were designed within exons 1 and 2 to amplify putative transcripts. The observed bands of the expected size 180 bp were sequenced and confirmed to correspond to SPANX-N genes. The strongest expression of SPANX-N genes was observed in the normal testis (lanes 12 and 17) and in the LOXIMV1 melanoma cell line (lanes 13 and 18). Lanes 2–11 correspond to normal and tumor pairs of breast, cervix, prostate, lung and ovary; lanes 19–21 correspond to 938MEL, 888MEL, SKMEL28 melanoma cell lines; lane 22 corresponds to the SKOV3 ovarian cell line; lanes 1, 15, 16, 24 – ladder; lanes 14 and 23 - water. The cDNA templates used were normalized using actin, as shown at the bottom of the panel. (b) Western blot analysis of lysates from normal tissues using an anti-EQPT antibody. Lane 1 - lung, lane 2 - testis, lane 3 - placenta and lane 4 – prostate. Lanes 5–9: full-size SPANX-N proteins expressed in the pET-11d in Bl21 cells. The mobility of SPANX-N proteins produced in E. coli cells is 13 kDa for SPANX-N1, 27 kDa for SPANX-N2, 23 kDa for SPANX-N3, 17 kDa for SPANX-N4 and 13 kDa for SPANX-N5.

Article Snippet: Interestingly, 92% of the sequenced RT-PCR products from different tumors and cancer cell lines corresponded to SPANX-N1 while only 3.3% of the clones from normal tissues corresponded to SPANX-N1 (mostly in testis), suggesting preferable expression of this gene in malignant tissues ( ).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Produced