FIGURE 5.

Rescue of pathologic phenotypes in edited N98S motor neurons. (A) Schematic of allelic series of iPSCs differentiated into healthy (continuous axon) or diseased (dashed axon) i³LMNs. Colored nuclei represent NEFL genotypes as described in Figure 3. Cytoplasmic shading indicates differential genetic backgrounds, dark gray cells are isogenic. (B) Representative images of day-7 i³LMNs stained with anti-NF-L (green) and anti-HB9 (blue) antibodies. Scale bars = 25 μm. (C) Quantification of NF-L relative fluorescence intensity (R.F.I.) in HB9+ cell bodies using automated image analysis pipeline. Data points represent mean values from independent differentiations, bars indicate mean of six biological replicates ± S.E.M. (D) ELISA for NF-L protein in media from day-14 i³LMNs. NF-L levels were normalized to neurite density measurements to correct for well-to-well variability in cell seeding (Supplementary Figure 8). Individual data points shown, bars indicate mean of five biological replicates ± S.E.M. For all graphs ****p < 0.0001 by one-way ANOVA with Šídák’s test for multiple comparisons.