Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Dec 21;110(24):4074–4089.e6. doi: 10.1016/j.neuron.2022.12.005

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2022 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

PlexinD1 fl/fl^Tie2:Cre embryos present increased spinal cord vascularization and altered MN maturation defects at E18.5

(A) Immunostaining for BVs (IsoB4⁺) in E18.5 spinal cords from PlexinD1 fl/fl and PlexinD1 fl/fl^Tie2:Cre embryos. Insets show higher magnification of one side of the ventral spinal cord. Scale bars, 100 μm.

(B) Quantification of BV density in the ventral spinal cord of PlexinD1 fl/fl and PlexinD1 fl/fl^Tie2:Cre embryos, normalized to control littermates at E18.5. n = 7 PlexinD1 fl/fl, n = 6 PlexinD1 fl/fl^Tie2:Cre, from two independent litters; parametric distribution, two-tailed unpaired Student’s t test. ∗p < 0.05.

(C and D) Representative images of RNAscope for Chat co-stained with HB9 (C) and respective quantification of the number of Chat dots per HB9⁺ nucleus (D) (normalized to control littermates). Scale bars, 25 μm. n = 5 PlexinD1 fl/fl, n = 4 PlexinD1 fl/fl^Tie2:Cre, from two independent litters; parametric distribution, two-tailed unpaired Student’s t test. ∗p < 0.05.

(E) Representative image of ChAT immunostaining in MNs from the medial motor column (MMC) and lateral motor column (LMC) of PlexinD1 fl/fl and PlexinD1 fl/fl^Tie2:Cre embryos at E18.5. Scale bars, 100 μm.

(F) Quantification of ChAT fluorescence intensity in PlexinD1 fl/fl and PlexinD1 fl/fl^Tie2:Cre embryos at E18.5. Data normalized to control. n = 6 PlexinD1 fl/fl, n = 6 PlexinD1 fl/fl^Tie2:Cre, from two independent litters; parametric distribution, two-tailed unpaired Student’s t test. ∗p < 0.05.

(G) Scheme representing the innervation of diaphragm by the phrenic nerve. Inset: location where pictures shown in (H) were taken.

(H and I) Representative images (H) of the diaphragm of PlexinD1 fl/fl and PlexinD1 fl/fl^Tie2:Cre embryos at E18.5 stained to show the phrenic nerve and its branching (Neurofilament M + Synapthophysin), AchR⁺ clusters (α-bungaratoxin); (I) representation of the output result of image analysis for branching points and length (using the neurofilament + synapthophysin staining). Scale bars, 100 μm.

(J) Quantification of total length of phrenic nerve branching in the diaphragm muscle at E18.5. Data normalized to control. n = 4 PlexinD1 fl/fl, n = 4 PlexinD1 fl/fl^Tie2:Cre, from two independent litters; parametric distribution, two-tailed unpaired Student’s t test. ∗p < 0.05.

(K) Quantification of branching density of phrenic nerve at E18.5 in PlexinD1 fl/fl and PlexinD1 fl/fl^Tie2:Cre embryos. Data normalized to control littermates. n = 4 PlexinD1 fl/fl, n = 4 PlexinD1 fl/fl^Tie2:Cre, from two independent litters; parametric distribution, two-tailed unpaired Student’s t test. ∗p < 0.05.

All data shown as mean ± SD.