Piwil1
(A) and Piwil2
(B) mRNA is detected in primary cortical neurons at baseline; mRNA of both Piwil1 and Piwil2 significantly increases following 3 hours of KCl stimulation (n=4–6, Student’s t-test, *p<0.05, **p <0.01). Expression of Piwil1
(C) and Piwil2
(D) is significantly reduced in primary cortical neurons 4 days after treatment with the respective shRNA-expressing AAVs, compared to neurons treated with a non-targeting control. (n=4, Student’s t-test, **p <0.01, ***p <0.001). (E)
Ppp3r1 mRNA induction by KCl-induced depolarisation (20mM, 3h) in primary cortical neurons is inhibited by simultaneous knockdown of Piwil1 and Piwil2 (n=3–5, one way ANOVA, F (3, 10) = 8.101, **p <0.01). (F) Simultaneous knockdown of Piwil1 and Piwil2 eliminates activity-induced ING1 occupancy at the Ppp3r1 upstream regulatory region under KCl-induced depolarisation (n=3–4, one way ANOVA, F (3, 9) = 4.556, *p <0.05). Con = scrambled non-targeting shRNA. Data represent mean ± SEM.