Supplementary Figure 2: Genome-editing to produce SH-SY5Y cells expressing TDP-43^N352S from both endogenous alleles.

(a) Histogram representation of the AAT codon (Asparagine) in position 352 of wild-type SH-SY5Y cells and its replacement by CRISPR-Cas9 to an AGT codon (Serine) in both endogenous TDP-43 alleles, as determined by sequencing in 3 independent biological samples (b) Nuclear to cytoplasmic TDP-43 ratios in SH-SY5Y^WT/WT (n = 34 cells, mean = 4.13) and SH-SY5Y^N352S/N352S (n = 31 cells, mean = 3.82) lines are plotted by using fluorescence intensity quantification. 3 biologically independent experiments were quantified. Statistical analysis was done using two-tailed t-test, P = 0.1427, SEM. (c) Alternative splicing alterations linked to TDP-43 loss of function ⁹ were tested in SH-SY5Y^WT/WT (black bars) and SH-SY5Y^N352S/N352S (red bars) lines. Representative images from 2% acrylamide gels are shown and inclusion/skipping band intensity ratios were plotted to the right (n = 5 independent biological experiments, two-tailed t-test, SEM). Mean values are represented for RWDD1 (2.08, P = 0.0002), CACNA1C (1.64, P = 0.00001), and ZNF569 (1.93, P = 0.003). **P < 0.01, ***P < 0.001. For uncropped gel images, see Supplementary Fig. 11. (d) Immunoblotting of stathmin-2 in SH-SY5Y^WT/WT and SH-SY5Y^N352S/N352S lines. α-Tubulin served as a loading control. Experiment was reproduced 3 times independently with similar results. For uncropped blot, see Supplementary Fig. 11.