Figure 6. Protein aggregates caused by... | Rockefeller University Press

Figure 6.

$Protein aggregates caused by SETX depletion are reduced by human RNaseH1 overexpression or transcription inhibition. (A) Detergent-resistant aggregates were isolated in U2OS cells with shRNA-mediated depletion of SETX, and induction of human wild-type RNaseH1 expression as indicated. PSMD2, TDP-43, and PSMD8 in lysate and aggregate fractions are shown by Western blotting. (B) Three replicates of A were performed and quantified; levels of PSMD2, TDP-43, and PSMD8 in aggregate fractions normalized by lysate levels are shown relative to control cells. (C) Aggregation assays as in A with SETX knock-out and human wild-type RNaseH1 overexpression as indicated. (D) Three replicates of C were performed and quantified; levels of PSMD2, TDP-43, and PSMD8 in aggregate fractions normalized by lysate levels are shown relative to control cells. (E) Aggregation assays as in A with control or SETX shRNA and DRB (20 μM) added as indicated. (F) Three replicates of E were performed and quantified; levels of PSMD2, TDP-43, and PSMD8 in aggregate fractions normalized by lysate levels are shown relative to control cells. All P values are derived using a two-tailed t test assuming unequal variance, using three biological replicates (n = 3). Error bars indicate standard error. *, **, ***, and **** indicate P < 0.05, 0.01, 0.001, and 0.0001, respectively; ns = not significant. Source data are available for this figure: SourceData F6.$

Protein aggregates caused by SETX depletion are reduced by human RNaseH1 overexpression or transcription inhibition. (A) Detergent-resistant aggregates were isolated in U2OS cells with shRNA-mediated depletion of SETX, and induction of human wild-type RNaseH1 expression as indicated. PSMD2, TDP-43, and PSMD8 in lysate and aggregate fractions are shown by Western blotting. (B) Three replicates of A were performed and quantified; levels of PSMD2, TDP-43, and PSMD8 in aggregate fractions normalized by lysate levels are shown relative to control cells. (C) Aggregation assays as in A with SETX knock-out and human wild-type RNaseH1 overexpression as indicated. (D) Three replicates of C were performed and quantified; levels of PSMD2, TDP-43, and PSMD8 in aggregate fractions normalized by lysate levels are shown relative to control cells. (E) Aggregation assays as in A with control or SETX shRNA and DRB (20 μM) added as indicated. (F) Three replicates of E were performed and quantified; levels of PSMD2, TDP-43, and PSMD8 in aggregate fractions normalized by lysate levels are shown relative to control cells. All P values are derived using a two-tailed t test assuming unequal variance, using three biological replicates (n = 3). Error bars indicate standard error. *, **, ***, and **** indicate P < 0.05, 0.01, 0.001, and 0.0001, respectively; ns = not significant. Source data are available for this figure: SourceData F6.

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