RT-PCR analysis of SPANX-N expression in normal and tumor tissues and cell lines

*SKOV3- ovarian; HeLa-cervix. Each RT-PCR product was cloned in TA vector. From 5 to 40 individual E. coli clones obtained for each sample were analyzed by sequencing. Cloning and sequencing of RT-PCR products determined the gene-specific transcripts in each tissue or cell line. 95 of 103 sequenced clones from different tumor tissues and cancer cell lines corresponded to SPANX-N1. Four of 120 sequenced clones from normal tissues corresponded to SPANX-N1. Expression of SPANX-N2 and SPANX-N3 proteins in lung, testis, placenta and prostate was confirmed by Western blot (Figure 6b).