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

. 2021 Nov 25;49(21):12284–12305. doi: 10.1093/nar/gkab1120

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

© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 4. — The hnRNP R-ΔN isoform is sufficient to support axon growth in primary motoneurons. (A) Western blot analysis of lysates from Hnrnpr⁺^/+ and Hnrnpr^tm1a/tm1a motoneurons cultured for 7 DIV, probed with an antibody against the C-terminus of hnRNP R. Gapdh was used as loading control. (B) Representative images of primary motoneurons cultured for 7 DIV and immunostained with antibodies against the C-terminal or N-terminal domain of hnRNP R, and with an antibody against β-Tubulin. Scale bars: 5 μm. (C) Quantification of mean fluorescence intensity of the hnRNP R immunosignal obtained with the C-terminal-specific antibody in motoneurons shown in (B). Data are mean ± SD (n = 3 independent experiments; N = 30 motoneurons each for Hnrnpr⁺^/+ and Hnrnpr^tm1a/tm1a). Statistical analysis was performed using two-way ANOVA followed by Bonferroni post-hoc test; n.s., not significant. (D) Quantification of the survival of motoneurons at 7 DIV as percentage of the number of motoneurons at 1 DIV, in the presence or absence of BDNF. Data are mean ± SD (n = 3 independent experiments; motoneurons were from N = 4 embryos each for Hnrnpr⁺^/+ and Hnrnpr^tm1a/tm1a). Statistical analysis was performed using two-way ANOVA followed by Bonferroni post-hoc test; n.s., not significant. (E) Box-and-Whisker plots of the axon lengths of motoneurons cultured for 7 DIV. Data are mean ± SD (n = 3 independent experiments; N = 461 motoneurons for Hnrnpr⁺^/+ and N = 502 motoneurons for Hnrnpr^tm1a/tm1a). Statistical analysis was performed using the Mann–Whitney test; n.s., not significant. (F) Box-and-Whisker plots of growth cone sizes of motoneurons cultured for 7 DIV. Data are mean ± SD (n = 3 independent experiments; N = 73 growth cones for Hnrnpr⁺^/+ and N = 86 growth cones for Hnrnpr^tm1a/tm1a). Statistical analysis was performed using the Mann–Whitney test; n.s., not significant. (G) Representative images showing 7SK RNA labeling by in situ hybridization and immunostaining for hnRNP R in motoneurons derived from Hnrnpr⁺^/+ and Hnrnpr^tm1a/tm1a mice. Arrowheads show 7SK-positive punctae. Scale bars: 5 μm. (H) Box-and-Whisker plots of the number of 7SK-positive punctae in axons of motoneurons cultured for 6 DIV. Data are mean ± SD (n = 3 independent experiments; N = 48 axons for Hnrnpr⁺^/+ and N = 55 axons for Hnrnpr^tm1a/tm1a). Statistical analysis was performed using the Mann–Whitney test; n.s., not significant.