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. 2008 Nov;22(11):3846–3852. doi: 10.1096/fj.08-110890

Figure 2.

Figure 2.

Allele-specific silencing of GNE in sialuria fibroblasts by siRNA. A) Location of the primer-probe assays used for gene expression analysis by real-time PCR in the GNE gene. Assays A and B, located over GNE exon-exon boundaries (exon 4–5 and exon 5–6, respectively), were employed for determination of total GNE expression. Assay C, located in exon 5 over the GNE mutation of the sialuria patient, was used for allele-specific expression analysis of the mutated (c.797A) and the wild-type (c.797G) allele. B) Real-time PCR analysis of total GNE RNA expression in sialuria fibroblasts 48 h after mock (no siRNA), siRNA-mut, and siRNA-wt transfections. C) Standard curve produced by mixing allelic quantities of mutated allele A vs. wild-type allele G, which was then used for determination of allelic expressions in patient cells. D) Allelic expressions of GNE [wild-type G-allele (white) vs. mutant A-allele (black)] in sialuria cells 48 h after mock (no siRNA), si-RNA-mut, and siRNA-wt transfections.