Nrg1 Intracellular Signaling Is Neuroprotective upon Stroke

Figure 2

Generation and validation of viral vectors for Nrg1 expression. (a) Schematic representation of the constructs for the expression of Nrg1-ICD (including the nuclear localization signal (NLS)) and Nrg1-FL both fused to GFP. The schema of the vector for the production of AAV particles to express Nrg1 under human Synapsin promoter is depicted on the right. CRD: cysteine-rich domain; EGF: epithelial growth factor domain; TM: transmembrane domain; ITR: inverted terminal repeat; ORF: open reading frame; hGH PA: human growth hormone polyadenylation signal. (b–j) Immunofluorescent labeling of primary cortical neurons infected to express GFP, Nrg1-ICD, or Nrg1-FL and fixed at D14. The neurons were labeled with DAPI and microtubule-associated protein 2 (MAP2) to visualize neuronal dendrites. The boxed area in (b, e, h) depicts the area magnified in (d, g, j). The dotted lines delimit the nuclei to highlight the specific localization of Nrg1 constructs. Scale bar, 50 μm.