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. 2008 Nov 27;3(11):e3823. doi: 10.1371/journal.pone.0003823

Figure 3. Prdm14 is overexpressed in Evi32 tumors.

Figure 3

3a) Q RT-PCR was performed on cDNA prepared from total RNA samples from AKXD tumors with and without insertions at Evi32 and control tissue obtained from 8 week old C57BL/6J mice. As before, expression levels were normalized to 18S expression then quantified. This analysis revealed that Prdm14 expression was upregulated in all Evi32 tumors. The expression levels of Prdm14 in Evi32 tumors varied between 30 and 50 fold more than in control spleen. 3b) Northern analysis of Prdm14 was performed using total RNA obtained from Evi32 tumors and control tissue obtained from 8 week old C57BL/6J mice. The probe to Prdm14 was nearly the full length coding sequence amplified by PCR. Sizes were estimated using 18S and 26S migration distances. This analysis shows specific upregulation of Prdm14 in all Evi32 tumors. 3c) RT-PCR was performed on Evi32 tumors using a viral 3′ LTR specific primer coupled with a Prdm14 second exon primer. cDNA obtained from the 27-001 produced a prominent band consistent with a viral fusion transcript. 3d) The amplicon from 27-001 was sequenced, and viral elements were found to be linked to the second exon of Prdm14 which contains the beginning of the coding sequence of Prdm14. SP = Spleen, LN = Lymph Node, Th = Thymus.