Fig. 3

SRSF6 levels modulate incomplete splicing of Htt. a–e Overexpression of SRSF6 increases the amount of incomplete splicing. Mouse Srsf6 (a) and human SRSF6 (b), respectively, were over-expressed to the same extent in the CAG₇ and CAG₁₀₀ cell lines. c Western blot data confirmed the over-expression of mouse and human SRSF6 for both CAG repeat lengths. - = mock; m = mouse SRSF6; h = human SRSF6. Uncropped blots can be found in Supplementary Fig. 8A. d Spliced exon 1–exon 2 transcripts were not significantly changed due to the overexpression of mouse or human SRSF6. e Intron 1 containing transcripts were increased to statistically significantly levels when human SRSF6 was over-expressed. Individual data points and the mean ± s.e.m. are shown. n = 8 independent experiments/CAG-length; two-way ANOVA with Tukey post hoc. Effect of SRSF6 overexpression for a given CAG-length: ^###p < 0.001. Effects due to CAG-length for a given treatment: ***p < 0.001. f–i Knock-down of SRSF6 by siRNA treatment (s12740, ThermoFisher) decreased the amount of incomplete splicing. SRSF6 levels were decreased by siRNA treatment on transcript (f) and protein (g) levels for both CAG repeat lengths. scr = scramble; si = siRNA treatment. Uncropped blots can be found in Supplementary Fig. 8B. h Spliced exon 1–exon 2 transcripts were not changed due to the knock-down of SRSF6. i There was a statistically significant reduction in the amount of incompletely spliced minigene in the CAG₁₀₀ line. Individual data points and the mean ± s.e.m. are shown. n = 3–6 independent experiments/CAG-length; two-way ANOVA with Tukey post-hoc. Treatment for a given CAG-length: *p < 0.05, **p < 0.01, ***p < 0.001. Treatment x CAG-length ^###p < 0.001