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

Figure 4

The p.G256S and p.R174H LFNG substitutions do not lead to aberrant protein processing. (a) Western blot of cell lysate from LFNG-HA transiently transfected NIH3T3 cells. Cell lysates were run on a 10% SDS gel, and membranes were incubated with anti-HA primary and anti-rabbit IgG secondary antibodies, or anti-actin primary and anti-mouse IgG secondary antibodies. (b, c) Densitometry analysis of pre-pro-LFNG-HA (b) or processed (c) LFNG-HA bands. Band intensities were normalized to actin. One-way ANOVA indicated a statistically significant difference between all conditions in both pre-pro-LFNG-HA (, ) and processed LFNG-HA (, ). Bonferroni-adjusted () post hoc analyses indicated that the pre-pro-LFNG-HA p.F188L bands were significantly more intense than WT, and the processed LFNG-HA p.F188L bands were significantly less intense than WT. No other conditions were significantly different from WT. Values are of three independent experiments () plated in triplicate. .

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