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

. 2023 Sep 22;14:5898. doi: 10.1038/s41467-023-41603-0

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

© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 5 — a Experimental setup for LPS-stimulated motor neuron (MN)-microglia (MG) co-cultures. b Percentage of MNs/MG in LPS-stimulated co-cultures of healthy control (HC) MNs and HC, isogenic control (IC), and C9orf72 mutant (C9) MG showing a ~ 30% microglia content (n = 3 microglial lines, n = 4 differentiations (C9-HC comparison), n = 1 microglial line, n = 5 differentiations (IC-C9-2 comparison)). c Exemplar Western blot against IBA1, ChAT, and MMP9 in LPS-stimulated co-cultures of HC MNs with MG. d Quantification of blot shown in c demonstrating pro-MMP9 and active MMP9 normalized to the housekeeping gene b-actin are significantly increased in LPS-stimulated co-cultures of HC MNs with C9 MG compared with controls (n = 3 microglial lines (C9-HC comparison), n = 1 microglial line (C9-2-IC comparison), n = 3 differentiations). e Exemplar images of apoptosis marker cleaved caspase 3 (CC3) in HC MNs in LPS-primed co-cultures with MG plus MMP9 inhibitor 1 (3 μM, MMP9i) or DMSO as vehicle control. Scale bars: 25 μm. f Quantification showing significantly increased relative fold change in CC3 expression in LPS-primed co-cultures with C9 MG compared with HC MG. Administration of MMP9i ameliorates microglial neurotoxicity. The C9-2-IC comparison shows the same pattern but does not reach statistical significance (n = 3 microglial lines, n = 4 differentiations (C9-HC comparison, n = 1 microglial line, n = 4 differentiations (IC-C9-2 comparison)). g Relative fold change in LPS-stimulated co-culture supernatant for selected targets using a cytokine array. Supernatant samples from LPS-primed co-cultures of HC MNs with C9 MG showing a dysregulated cytokine profile compared with HC MG, which is ameliorated by MMP9i treatment (pooled samples from n = 3 microglial lines with n = 3 differentiations each). h ELISA quantification of DPP4 in co-culture supernatants from LPS-stimulated co-cultures of HC MNs and MG showing significantly increased DPP4 levels, which is significantly reduced after MMP9i treatment (n = 3 microglial lines (C9-HC comparison), n = 1 microglial line (IC-C9-2 comparison), n = 3 differentiations). Single data points and means ± SEM. *P < 0.05; **P < 0.01; ****P < 0.0001; ns: not significant. Two-tailed unpaired t-test (d) and one-way ANOVA with Tukey’s post-hoc test (f, h). The graphics for panel a were created using Biorender.com. Source data are provided as a Source Data file.