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Fig. 5. Intrinsic Sema3A cell-autonomously modulates the Nrp1 level at the
growth cone surface. (A) In situ hybridization of Nrp1
mRNA (NP1) on spinal transverse sections from Sema3AiresEGFP and EGFP
embryos. Brackets indicate the electroporated side. Scale bar: 100 µm.
(B,C) Nrp1 and NgCAM immunostaining of EGFP+ and
EGFP- growth cones of Sema3AiresEGFP and ctrlEGFP explants, with
membrane permeabilization (B), and without permeabilization (C). Scale bars:
10 µm. (D-G) Histograms showing fluorescence intensities per unit of
surface at the growth cone. Fluorescence intensities were normalized to
EGFP+ conditions in D-F. Fluorescence intensities of
pShScramble-EGFP+ and pShSema3A-EGFP+ growth cones were
normalized to the pShScramble-EGFP- or pShSema3A-EGFP-
conditions, respectively (G). **, P<0.01;
***, P<0.001 with Mann-Whitney test. The number of
growth cones analyzed is indicated for each condition. (H) Nrp1 and
NgCAM immunostaining of EGFP+ and EGFP- growth cones of
pShScramble-EGFP and pShSema3A-EGFP explants without permeabilization.
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