Functional Characterization of Novel Lunatic Fringe Variants in Spondylocostal Dysostosis Type-III with Scoliosis

Figure 3

The p.G256S and p.R174H LFNG substitutions are null and hypomorphic, respectively. (a) Western blot of anti-FLAG immunoprecipitated 3XFLAG-LFNG protein. 3XFLAG-LFNG was purified with anti-FLAG agarose resin and disassociated with SDS buffer, and 20 μL of each sample was run on a 10% SDS gel. Protein was detected with anti-FLAG primary antibody and HRP-linked anti-mouse IgG secondary. (b) GlcNAc transferase activity of LFNG variant protein isolated from media. One-way ANOVA indicated a significant difference between groups (, ), and Bonferroni-adjusted () post hoc analyses indicated that WT was significantly different from p.R174H, p.G256S, and E.V. p.R174H was significantly more intense than p.G256S and EV. There was no statistically significant difference between p.G256S and EV (). Values are of three independent experiments () plated in triplicate. and .

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