Figure 10.
G-rich segment in intron 5 of SIGLEC14 pre-mRNA suppresses splicing of the intron. Differentials in the threshold cycle (ΔCT) between spliced (membrane-bound) versus unspliced (soluble) SIGLEC14 mRNA isoforms in 293T cells transfected with Sig14+I5/pcDNA (mini-gene construct with intron 5) or with Sig14+I5Δ27/pcDNA (mini-gene construct with intron 5 but lacking 27-nucleotide G-rich segment) were analyzed by quantitative RT-PCR. Each data point represents a mean of two to three technical replicates (i.e. two to three wells each of 293T cells were transfected with either Sig14+I5/pcDNA or Sig14+I5Δ27/pcDNA in parallel;, ΔCT for mRNA isoforms were analyzed by qRT-PCR for each well, and mean ΔCT was calculated for each construct). The experiments were repeated seven times. The dataset was analyzed by paired t test. **, p < 0.01.