FIGURE 2.

Astrocyte derived EV‐IL‐1β promotes APP translation through hnRNP C. Primary neurons were treated with vehicle (control), astrocyte derived EV‐CR (ADEV‐CR) or astrocyte derived EV‐IL‐1β (ADEV‐IL‐1β) (50 EVs per neuron) for 6–24 h as indicated. (a) Representative Western blot showing APP and BACE1 detected from neuronal lysates 24 h following the indicated treatment conditions. Bar graphs show densitometric quantitation of APP and BACE1 following the indicated treatments and time points. Data are mean ± SEM of n = 3 independent experiments per condition. *P < 0.05, **P < 0.01, one‐way ANOVA with Tukey post‐hoc comparisons. (b) Bar graphs show quantitative RT‐qPCR of APP and BACE1 transcripts following the indicated treatment conditions. Data are mean ± SEM of n = 3–6 independent experiments per condition. (c) APP mRNA binding to hnRNP C was measured by RT‐qPCR. Data are mean ± SEM of n = 4–6 experiments per condition. *P< 0.05, one‐way ANOVA with Tukey post‐hoc comparisons. (d) Representative Western blot showing total hnRNPC and phosphorylated hnRNPC detected using Ab‐Dynabeads to isolate hnRNPC followed by immunoblot using a phosphorylation antibody. Data are mean ± SEM of n = 3 experiments per condition. *P < 0.05, one‐way ANOVA with Tukey post‐hoc comparisons. (e) hnRNP C mRNA measured by RT‐qPCR and (f) representative immunoblot and densitometric quantitation of hnRNP C protein levels following the indicated treatments Data are mean ± SEM of n = 3‐6 independent experiment per condition. *P < 0.05, **P < 0.01, one‐way ANOVA with Tukey post‐hoc comparisons