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

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© 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/.

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Fig. 3 — a ELISA quantification of MMP9 in supernatants from unstimulated and LPS-stimulated healthy control (HC), isogenic control (IC), and C9orf72 mutant (C9) microglia in monoculture (pMGL) shows significantly increased MMP9 release in the C9-HC (n = 3 lines per condition, n = 5 differentiations each) and IC-C9-2 pMGL comparisons (n = 1 line per condition, n = 6 differentiations each). b Fluorometic activity assay for active MMP9 in supernatants from unstimulated and LPS-stimulated HC, IC, and C9 pMGL shows significantly increased MMP9 activity in the C9-HC (n = 3 lines per condition, n = 5 differentiations each) and IC-C9-2 pMGL comparison (n = 1 line per condition, n = 6 differentiations each). c RT-qPCR for MMP9 expression in unstimulated and LPS-stimulated HC, IC, and C9 pMGL normalized to the housekeeping gene TBP showing significantly increased MMP9 expression in the C9-HC (n = 3 lines per condition, n = 3 differentiations each) and numeric increase in IC-C9-2 pMGL comparison (n = 1 line per condition, n = 3 differentiations each). d Top: exemplar Western blot against the pro-form and active form of MMP9 in unstimulated and LPS-stimulated HC, IC, and C9 pMGL. Bottom: quantification shows significantly increased expression of pro-MMP9 in the C9-HC pMGL comparison and a numeric increase in the IC-C9-2 pMGL comparison. Active MMP9 is significantly increased in both comparisons. Both normalized to the housekeeping gene b-actin (n = 3 lines per condition for C9-HC comparison, n = 1 line for the IC-C9-2 comparison, n = 4 differentiations each). Single data points and means ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. One-way ANOVA with Tukey’s post-hoc test (a–d). Source data are provided as a Source Data file.