Growth Cone MKK7 mRNA Targeting Regulates MAP1b-Dependent Microtubule Bundling to Control Neurite Elongation
Figure 8
Validation of neurite-localized JNK network in hippocampal neurons.
E18 hippocampal neurons were plated on poly-D-lysine–coated coverslips. Cells were fixed at DIV 1. (A) Confocal fluorescence micrographs of MKK7 mRNA FISH using antisense probe. Top panels: fluorescence signal in ibw contrast. Bottom panels: composite image of FISH and phalloidin immunostaining. Arrowheads point to MKK7 mRNA. FISH probe was directed against the 3′-UTR2 region. (B) Sense MKK7 mRNA FISH control. Fluorescence intensities were scaled as in (A). (C) Confocal fluorescence micrographs of neurons immunostained for different components. Images are color coded for fluorescence intensity so that warm and cold colors represent high and low signal intensity, respectively. F-actin panels are shown in ibw contrast. (D) Averaged fluorescence intensity profiles along chosen neurites. Red dotted lines represent the soma/neurite interface whereas blue dotted line represents neurite/growth cone interface. Fluorescence images were subjected to line scan analysis with a 70-µm long line. The mean ± SEM of normalized fluorescence intensity profiles from ten neurites are shown. Scale bars: 10 µm.