Bioinformatic screening
Animal and human samples
Cell culture and transfections
Plasmids, shRNAs, and DNA cloning
Virus preparation
SV exo-endocytosis evaluation and calcium imaging
Neurite elongation assay
In utero electroporation
Single-molecule RNA in situ hybridization and immunofluorescence
Quantitative real-time PCR
Library preparation and sequencing
RNA-sequencing analysis
ChIRP DNA analysis
GO and STRING analyses
5′-RACE PCR
ChIRP and RNA interactome analysis
MS identification of protein interactors following ChIRP/RNA interactome analysis
RNA immunoprecipitation
Western blot
Immunocytochemistry
Statistical analysis and graph construction
For the pharmacological modulation of neuronal activity, we used 1 μM TTX (Na+ Channel blocker; Cayman Chemical, USA), 20 μM bicuculline methobromide (GABAA receptor antagonist; Tocris Bioscience, Cambridge, UK), 100 μM 4-AP (K+ channel blocker; Tocris Bioscience, Cambridge, UK), 10 μM CNQX (AMPA/kainate receptor antagonist; Tocris Bioscience, Cambridge, UK), and 50 μM D-AP5 (N-methyl-d-aspartate receptor antagonist; Tocris Bioscience, Cambridge, UK). A total of >200 neurons from three different cultures were blindly analyzed for each condition.