Fig 2: miR376c-Mediated Gene Silencing Is Reduced after iLTP(A) qRT-PCR showing miRNA expression levels following iLTP induction (normalized to U6 snRNA). Colored stars compare statistical significance to the ctrl; n = 4.(B) Luc-GABRA1 activity in neurons under control conditions or 90 min post-iLTP induction; n = 5.(C) As in (B) but for Luc-GABRG2; n = 5(D) Luc-GABRA1 activity in neurons expressing anti-miRCon or anti-miR376c, under control conditions or 90 min post-iLTP induction; n = 5.(E) As in (D), but for Luc-GABRG2; n = 5.(F) a1, ?2, and gephyrin levels 90 min post-iLTP induction in neurons expressing miRCon or miR376c; n = 6.(G) GFP-filled dendritic segments from miRNA overexpressing hippocampal neurons labeled with antibodies to surface GABAAR?2 and gephyrin or VGAT. Scale bar, 10 µm.(H) Quantification of cluster area and density from (E); n = 16–18 neurons per condition. Colored stars compare statistical significance with ctrl.(I) mIPSC traces from neurons infected with miRCon or miR376c. Graphs show mIPSC amplitude and frequency; n = 9–13 cells per condition.All values represent mean ± SEM. *p < 0.05 and **p < 0.01, one-way ANOVA, Bonferroni post hoc test (A) and two-way ANOVA, Bonferroni post hoc test (B–F and I).

Fig 3: miR376c Silences GABRA1 and GABRG2(A) qRT-PCR measuring mRNA levels 90 min post-iLTP induction. Readings were normalized to U6 small nuclear RNA (snRNA) levels; n = 5.(B) Ago2 was immunoprecipitated equally from control neurons and from neurons 90 min post-iLTP induction. qRT-PCR measured mRNA bound to Ago2. Readings were normalized to total levels from (A); n = 5.(C) Schematic of the Luc-GABRA1 luciferase reporters. miR376c seed site is mutated in Luc-GABRA1Mut. Graph of Luc-GABRA1 activities in HEK293T cells co-expressing miR376c, miRCon, or no miRNA. Firefly activities were normalized to Renilla activity; n = 9.(D) As in (C) but for GABRG2; n = 10.(E) Levels of a1 and ?2 in neurons expressing miRCon or miR376c; n = 6. miRNA expression constructs contain a GFP reporter.(F) Levels of a1 and ?2 in neurons expressing anti-miRCon or anti-miR376c; n = 5.All values represent mean ± SEM. *p < 0.05 and **p < 0.01, t test (A, B, E, and F) and two-way ANOVA, Bonferroni post hoc test (C and D).