Fig. 7.

Overexpression of Nova in DRG neurons promotes the “mature” splicing pattern observed in CNS neurons. a Schematic illustrating overexpression of Nova1 in rat primary DRG neuronal culture using a Nova1-GFP-expressing lentivirus. A GFP-expressing lentivirus is used as a Mock control. b qRT-PCR of Nova1 mRNA expression level in DRG neurons transduced with Nova1-GFP-expressing or GFP-expressing lentiviruses. The average relative expression level normalized to Mock-transduced cells is shown. Error bars represent standard deviation (n = 3). c Immunostaining of primary rat DRG neurons transduced with Nova1-GFP-expressing or GFP-expressing lentiviruses. Nova1 protein has predominant nuclear localization. Scale bar, 50 µm. The insert shows the zoom-in view of the cell. d Changes in the alternative splicing of Nova target exons upon Nova1 overexpression in rat primary DRG neurons. For each exon, the exon inclusion level in WT and Nova2 KO cortex, and DRG sensory neurons, as quantified by RNA-seq, is shown in the barplot on the left. Error bars represent the standard deviation (n ≥ 3). A representative gel image on the right shows inclusion of the alternative exon in Mock-transduced and Nova1-transduced cells detected using RT-PCR analysis (n = 3). The bands corresponding to the inclusion (top) and skipping of the exon (bottom) are indicated