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. Author manuscript; available in PMC: 2017 Mar 1.
Published in final edited form as: Mol Cell Neurosci. 2015 Dec 17;71:34–45. doi: 10.1016/j.mcn.2015.12.008

Figure 6. Over-expression of TDP-43 reduces miRNA regulation of EIF2C4/AGO4 stability.

Figure 6

(A) Schematic representation of protein expression when N-Flag-TDP-43 AinV15 cells containing an inducible tetracycline response element (Tet) are grown in the presence or absence of Dox for 5 days. (B) Total cell lysates were immunoblotted for the Flag tag to confirm the expression of Flag-TDP-43 in the presence or absence of Dox. Actin levels were assessed to ensure equal protein loading. (C) At day 3 of Dox induction, the cells were transiently transfected with a luciferase reporter fused to the 3’ UTR of EIF2C4/AGO4 (75-929 nt). The reporter harbors seed sequences recognized by three of the 5 selected DEmiRNAs, including miR-140-3p, miR-577, and Let-7e. (D) Relative luciferase activity was quantified in duplicate to determine the effect of endogenous miRNAs on luciferase reporter expression in the presence or absence of TDP-43 over-expression. Data are presented as the mean ± SEM of three independent experiments. ***P<0.001.