Figure 3.
Correlation of EGFP expression with Nav1.8 protein in DRG neurons from Scn10a-EGFP mice. A, Immunoreactivity (IR) of DRGs from an Scn10a-EGFP mouse to antibodies against Nav1.8 protein (NeuroMab). Phase-contrast (left), EGFP (middle), and red fluorescence (right) images of immunostained DRGs. Top, IHC of sectioned DRG tissue. Bottom, ICC of dissociated DRG neurons. White arrowheads indicate EGFP-positive DRG neurons lacking Nav1.8-IR. Yellow arrowheads indicate Nav1.8-IR DRG neurons lacking EGFP expression. Scale bar, 50 μm. B, Scatterplot from a representative ICC experiment. Green and red fluorescence intensity represents transgene (EGFP) expression and Nav1.8-IR, respectively, of DRG neurons from Scn10a-EGFP mice. ROIs were automatically selected around dissociated DRG neurons in the phase-contrast image, and fluorescence intensity was measured from subsequent images collected from green and red channels. Each dot indicates an individual cell. Dashed lines indicate the boundary between groups determined by staining in negative controls (secondary antibody alone) and visual confirmation.