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. 2011 Aug 24;6(8):e23878. doi: 10.1371/journal.pone.0023878

Figure 2. Expression of splice variants of ZnT5 detected by RT-PCR.

Figure 2

(A) Variant B is a genuine transcript of the ZnT5 gene. Product corresponding to a full-length ZnT5 variant B mRNA (1569 bp, lane 1) was generated by RT-PCR from total RNA extracted from human intestinal Caco-2 cells grown for 24 hours under standard conditions, which does not promote polarisation. Identity to the expected product was confirmed by sequencing. Molecular weight markers with sizes indicated shown (lane M). (B) Tissue-specific RT-PCR carried out using primers designed to anneal to the unique 3′ end of variant A (exons 13–17) and variant B (exons 13–14 3′) in a multiple tissue RNA panel (Ambion) revealed ubiquitous expression of both transcripts. Identity to the expected products was confirmed by sequencing. Negative control RT-PCR reactions identical to those yielding the products shown except for the omission of Moloney murine leukemia virus reverse transcriptase generated no products. Control reactions for all samples using primers to GAPDH [11] were including to control for input (results not shown).