Figure 4.
The WRC is expressed in spinal cord commissural neurons
(A) The mean mRNA expression levels (±SEM) of the WRC components in dissociated rat commissural neurons (n = 3 independent experiments) from RNA sequencing of dissociated cultured embryonic rat commissural neurons (GSE268644; from Makihara et al.9). The expression level of Boc is included for comparison.
(B) Immunostaining of dissociated E13.5 rat commissural neurons for Cyfip2, Cyfip1/2, Abi1, and Nckap1. Two examples are shown for each antibody. F-actin was labeled with phalloidin. Cyan arrowheads indicate Cyfip2, Cyfip1/2, Abi1, or Nckap1 at the leading edge of the growth cone. Scale bar: 10 μm.
(C) Schematic representation of the trajectories of commissural axons in a spinal cord cross-section of E11.5 mouse. Commissural neurons located in the dorsal spinal cord project axons that are guided ventrally toward the floor plate. Immunostaining of Cyfip1/2, Abi1, or Nckap1 with the commissural neuron marker Robo3 in E11.5 mouse spinal cord sections. (Upper row) Yellow arrows delimitate the commissural axon tract. Yellow arrowhead indicates the floor plate. Scale bar: 200 μm. (Lower row) A zoom in of the ventral spinal cord is below each corresponding image. Cyan arrowheads indicate some commissural axons. Scale bar: 100 μm. See also Figures S4–S7.