Figure 6.

TDP-43 overexpression promotes RNA instability in an exosome-dependent fashion. (A) Transcript half-life was estimated after ActD (5 μg/ml) treatment of HEK293 control and induced cells. Transcript levels at the indicated times were analyzed by qPCR. The relative levels of expression at the different time points were normalized according to the levels at time 0, (−) and (+) Tet induction (empty and filled circles, respectively). (B) Exosome components, PM/Scl-100 and DIS3, were simultaneously silenced in control and induced cells and their effect on TDP-43 expression levels was observed in western blot. (C) Quantification of endogenous TDP-43 protein levels under the different conditions was normalized against p84 levels. Error bars, s.e.m.; n=3 experiments for both panels. (D) mRNA expression levels of endogenous TDP-43 mRNA levels in −Tet and +Tet conditions. Both the transcript levels were normalized to the GAPDH mRNA levels and determined by real-time qPCR. The mRNA levels were quantified separately in triplicates and s.d. values obtained are shown on the bars.