Supplementary Figure 4: Incorporation of exon 2a in stathmin-2 mRNA is not followed by splicing of downstream exons.

(a) Representation of possible stathmin-2 splice variants resulted from loss of TDP-43. (b) Schematics of possible stathmin-2 RNAs with exons 1 and 2. (c) RT-PCR demonstrating reduced stathmin-2 mRNA after TDP-43 depletion without expression of an isoform containing exon 2a spliced to exon 2, diagram is shown to the right. Experiment was repeated independently 3 times with similar results. Unprocessed gel image is shown in Supplementary Fig. 11. (d) Conservation scheme of the polyadenylation signal embedded within stathmin-2 exon 2a. Thirteen primate species, mouse and rat genomic regions are represented. (e) qPCR analysis of stathmin-2 transcripts demonstrates reduced full-length stathmin-2 pre-mRNA in SH-SY5Y cells expressing mutant TDP-43 in comparison to wild type SH-SY5Y cells. Mean values of 2 independent biological experiments are plotted. Error bars represent SD. Expression of the nuclear non-coding RNA Xist was used as endogenous control.