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

. 2020 Nov 30;11:6131. doi: 10.1038/s41467-020-19914-3

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

© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 8 — a–f Representative images of the distal-most portion of axons from mCherry⁺ (a, b) or hM3Dq⁺ (c, d) DRGs stained for βIII-tubulin (green) and acetylated tubulin (red in a and c; standard fire scale pseudo-color in b and d). Images were taken 24 h after replating and CNO treatment. The distal-most 50 µm portion of axons growing from mCherry⁺ neurons and chemogenetically activated, hM3Dq⁺ neurons had a similar fluorescence intensity for βIII-tubulin (e). However, the ratio of acetylated tubulin to total tubulin (βIII-tubulin) was significantly smaller in the axons from CNO-activated, hM3Dq⁺ DRG neurons (f), indicating a decrease in stable, acetylated microtubules in axons extending from chemogenetically-activated neurons. g, h Cultures of mCherry⁺ or hM3Dq⁺ DRGs on ChABC-treated aggrecan spots were treated with CNO and either the HDAC inhibitor MS-275 or DMSO control. Chemogenetic activation alone decreased acetylated tubulin levels in the distal axon, compared to mCherry⁺ controls. Adding MS-275 to activated, hM3Dq⁺ DRGs increased the ratio of acetylated tubulin to total tubulin in the distal axon end to one that is comparable to mCherry⁺ (g) and abolished the improved ability of activated, hM3Dq⁺ neurons to cross the inhibitory rim of the aggrecan spot (h). N = 72 mCherry⁺ neurons and 75 hM3Dq⁺ neurons in e and f. N = 84 mCherry⁺+vehicle neurons, 88 hM3Dq⁺+vehicle neurons, 74 hM3Dq⁺+MS-275 neurons, and 67 mCherry⁺+MS-275 neurons in g. N = 24 aggrecan spots (6 coverslips)/group in h. Mean ± SEM. Two-tailed unpaired t-test in f and one-way ANOVA and post-hoc multiple comparisons testing using the two-stage step-up method of Benjamini, Krieger, and Yekutieli in g and h. *p < 0.05 (p = 0.0434 in f; mCherry+vehicle vs. hM3Dq+vehicle p = 0.0436, hM3Dq+vehicle vs. hM3Dq+MS-275 p = 0.0250 in g), **p < 0.01 (hM3Dq+vehicle vs. mCherry+MS-275 p = 0.0035 in g; hM3Dq+vehicle vs. hM3Dq+MS-275 p = 0.0071, hM3Dq+MS-275 vs. mCherry+MS-275 p = 0.0045 in h), ***p = 0.0002, ****p < 0.0001. Scale bar: 10 µm. Source data are provided as a Source Data file.