Figure 6.

Detailed characterization of hnRNPC binding sites near Tau exon 11. (a) E11-2 is divided into four sub-fragments, 150 nt each with 20 nt overlap. hnRNPC interacts with E11-2-1 and E11-2-2 as detected by in vitro RNA pull-down assay. The binding fragments are marked in red. (b) U-tract positions in E11-2-1 and E11-2-2 are highlighted in yellow. (c) Interaction between hnRNPC and the E11-2-1 sub-fragment with mutated U-tract motifs (by replacing the middle uridine with cytosine) separately and altogether. E11-2-1-1 m represents first U-tract was mutated; E11-2-1-2 m represents second U-tract was mutated; E11-2-1-12 m represents both U- tracts were mutated. (d) Interaction between hnRNPC and the E11-2-2 sub-fragment with mutated U-tract motifs following the same procedure described above. (e) Interaction between hnRNPC and E11-2 fragment is largely abolished with the mutations in first four U-tracts (shown as E11- 2-4Um) compared to mutations in all U-tracts. (f) First four U-tracts in E11-2 fragment (underlined in red in panel b) together with the U-tracts in E10-1 fragment were mutated in Tau minigene (shown as TM_E10-1 m&E11-2-4Um). The impact of hnRNPC on Tau exon 10 inclusion was mostly abolished when co-expressed with this mutated Tau minigene compared to full U- tracts mutated Tau minigene that described in Figure 5(e). Results shown are from three independent experiments.