Fig. 1

Validation of neuronal translatome enrichment in TRAP-RNA from Camk2a-NuTRAP mouse hippocampus. A Imaging of the hippocampal dentate gyrus demonstrated EGFP and mCherry co-expression in NeuN + cells. B TRAP-isolated hippocampal RNA from input, negative, and positive fractions were assessed by qPCR for enrichment and depletion of canonical marker genes for microglia, astrocytes, oligodendrocytes, and neurons. Mean relative gene expression ± SEM scaled to input for each gene. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by RM one-way ANOVA with Tukey’s multiple comparison test across fractions (n = 4/group). C RNA-seq was performed for all fractions (n = 4/group). Principal component analysis shows separation of the positive from input and negative fraction samples in the first component. D Cell-type marker gene lists were examined for fold change (Positive/Input) enrichment or depletion shows enrichment of neuronal markers and depletion of other cell-type markers in the positive fraction. E CIBERSORTx calculation of cell type composition of each fraction. The positive fraction is estimated to contain 100% neurons. F Genes with significant enrichment (2111) or depletion (2897) in the positive compared to input fraction were identified (FC >|1.25|, p < 0.05, Benjamini Hochberg multiple testing corrections). G–J Gene Ontology enrichment analysis and Ingenuity Pathway Analysis performed on significantly enriched or depleted genes (Positive/Input fraction) identified in E