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. 2024 Jun 8;12:90. doi: 10.1186/s40478-024-01807-x

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© The Author(s) 2024

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Mitophagosomes, mitolysosomes, autophagosomes and lysosomes as well as mitochondrial content analyses reveal defective mitophagy and autophagy processes in AD-D fibroblasts. a, c Representative images of Mit-RFP (red), LC3-GFP (a) or LAMP1-GFP (b) (green) signals in CTRL, AD-MCI and AD-D fibroblasts. The merged signal (yellow) reflects the colocalization of the red and green signals. Scale bars = 2 μm. b, d Quantitative graphs of the colocalization (Pearson’s coefficient) of Mit-RFP with LC3-GFP from CTRL (5 individuals, n = 42 cells), AD-MCI (6 patients, n = 49 cells), and AD-D (5 patients, n = 33 cells) (b) or of Mit-RFP with LAMP1-GFP from CTRL (5 individuals, n = 41 cells), AD-MCI (5 patients, n = 40 cells), and AD-D (5 patients, n = 40 cells) (d). Means ± SEMs *p < 0.05; **p < 0.01; ***p < 0.001; ns: not significant. e–j Representative images and masks of LC3-GFP (e) and LAMP1-GFP (h) signals in CTRL, AD-MCI and AD-D fibroblasts. Scale bar = 2 µm. f, i Quantitative graph of the number ± SEM of LC3-GFP puncta (f) and LAMP1 puncta (i). g, j Quantitative graph of the size ± SEM of LC3-GFP puncta (g) and LAMP1 puncta (j). The data were obtained from the CTRL group (5 individuals, n = 41 cells), AD-MCI group (6 patients, n = 49 cells) and AD-D group (5 patients, n = 40 cells). *p < 0.05; **p < 0.01; ***p < 0.001; ns: not significant. K–n SDS‒PAGE (k) and quantitative graphs of Parkin (l), TOMM20 (m), CPN10 (n), and CoxII (o) expression levels ± SEM obtained in mitochondria-enriched fractions of CTRL (n = 9), AD-MCI (n = 11) and AD-D (n = 9) fibroblasts. The data were normalized to those of CTRL fibroblasts. *p < 0.05; **p < 0.01; ns: not significant. p Heatmap of the correlation matrix computed linear regression (R²) between TOMM20, CPN10 and CoxII expression levels and the clinical Dementia Rating (Sum of Boxes) (CDR-SOB), the Mini Mental State Examination (MMSE) and the β-amyloid plaque burden assessed using the ¹¹C‐labelled Pittsburgh compound B (Global Cortical Index) (PiB-GCI) through positron emission tomography (PET-scan) at patient inclusion. A color scale of 1 corresponds to the maximum positive correlation value (blue), and a scale of − 1 corresponds to the maximum negative correlation value (red). *p < 0.05 for every pair of data sets