Fig. 6

Numa1 expression is post-transcriptionally regulated through its 3’UTR. A Numa1 levels measured by RT-qPCR from WT and Mov10 Deletion DIV4 hippocampal cultures treated with DRB. Numa 1 half-life in WT = 4.93 h and in Mov10 Deletion (DEL) = 3.54 h. B Luciferase expression (Renilla/Firefly luciferease) in Neuro2A WT and Mov10 knockout (KO) cells transfected with psiCHECK-2-Numa1−3’UTR and either no miR (NC) or miR-124, −133, −335, and −543. Data are shown as mean ± SEM. P-values were calculated using Mann–Whitney U test. **p-value < 0.01, ***p-value < 0.001, n = 11, N = 3. C AGO2-eCLIP reads in Numa1 from Immunoprecipitations (IP) from P0 WT brain (WT1 IP) (“Input” is total brain RNA) and two Mov10 Deletion P0 brains DEL1 IP and DEL2 IP. WT2 IP is from a previously published AGO2 eCLIP [5]. Peaks (in blue) are the input-normalized clusters identified by CLIPper (v2.0.1). The track height range for all tracks is 0–30. Data are visualized in IGV (v2.18.0) and the location of the MREs are indicated below. D Model of Numa1 3’UTR (NM_001403544.1) with AGO2 eCLIP sites identified in (C) and MOV10 CLIP sites identified in [34], miR-124, −133, −335, and −543 miRNA recognition sites, and predicted G-quadruplexes [41] and an in vitro G-quadruplex indicated [42]. “X” indicates that MOV10 blocks miR-543 MRE in Neuro2A while in brain (checkmark) MOV10 appears to facilitate AGO2 association