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. 2023 Jul 17;5(8):1290–1302. doi: 10.1038/s42255-023-00835-6

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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 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/.

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Fig. 3 — a, Free CI and CI-containing SCs (SC-CI) in WT and Cpt1a KO primary astrocytes, analysed by BNGE followed by immunoblotting against CI subunit NDUFA9. Data are mean ± s.e.m. P values are indicated (n = 3 biologically independent cell culture preparations; unpaired Student’s t-test, two-sided). b, Free CIII and CIII-containing SCs (SC-CIII) in WT and Cpt1a KO primary astrocytes, analysed by BNGE followed by immunoblotting against CIII subunit UQCRC2. Data are mean ± s.e.m. P values are indicated (n = 3 biologically independent cell culture preparations; unpaired Student’s t-test, two-sided). c, H₂O₂ production in WT and CPT1A KO astrocytes in primary culture. Data are mean ± s.e.m. P values are indicated (n = 6 independent cell culture preparations; unpaired Student’s t-test, two-sided). d, OCR analysis in WT and Cpt1a KO astrocytes in primary culture, either transfected with scrambled (control) or NDUFS1 siRNAs. Data are mean ± s.e.m. P values are indicated (n = 4 biologically independent cell culture preparations) (Supplementary Fig. 3f). e, Basal respiration in WT and CPT1A KO astrocytes in primary culture, either transfected with scrambled (control) or NDUFS1 siRNAs. Data are mean ± s.e.m. P values are indicated (n = 4 biologically independent cell culture preparations; two-way ANOVA followed by Tukey). f, H₂O₂ production by WT and CPTA1A KO astrocytes in primary culture, either transfected with scrambled (control) or NDUFS1 siRNAs. Data are mean ± s.e.m. P values are indicated (n = 3 biologically independent cell culture preparations; multiple unpaired Student’s t-test). g, Strategy used to assess the effect of Cpt1a KO astrocytes on WT or mCAT neurons in primary culture. Created with BioRender.com. h–l, Glutathione concentration (h), mitochondrial ROS (i), ∆ψ_m (j), apoptosis (k) and c-Fos and Arc mRNA abundances (l) in WT or mCAT-expressing transgenic neurons after coculture with WT or Cpt1a KO astrocytes; n = 3 (h), 4 (i), 4 (j), 4 (k, WT), 4 (k, mitoCAT) and 4 (l) biologically independent cell culture preparations; paired Student’s t-test, two-sided for simple comparisons and two-way ANOVA followed by Tukey for multiple comparisons.

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