EMBO/soda-nel huggingface dataset:
CCR5 deficiency impairs CD4+ T cell memory responses and antigenic sensitivity through increased ceramide synthesis (2020)
Figure 7

Figure 7. CCR5 controls TCR nanoclustering and Cer metabolism in human CD4+ cells. A. Analysis of TCR nanoclustering in lymphoblasts from healthy WT and ccr5Δ32 homozygous donors by EM. Top, representative small field image showing gold particle distribution in cell surface replicas of anti-CD3ε-labeled cells; bottom, quantification (mean ± SEM) of gold particles in clusters of the indicated size in the WT (gray bars; n = 5 cells, 17689 particles) and Δ32/Δ32 cells (light red; n = 4 cells, 16938 particles). Insets show the distribution between clusters of one, two, three, four or more than four particles, and statistical analysis.

B. Normalized SM, Cer and dhCer levels in lymphoblasts obtained as in A. A representative experiment is shown (n = 3 donors/genotype; n = 2 independent experiments)

C. Relative CerS2 mRNA levels in day 8 WT and ccr5Δ32 lymphoblasts. Each data point is the average of a technical triplicate from 3 donors in two independent experiments (n = 6)

Data information: Data are shown as mean ± SEM (B, C). * p <0.05, ** p <0.01, two-tailed unpaired Student's t-test. Scale bar, 50 nm.

EXPANDED VIEW FIGURE LEGENDS
Molecular diversity of diencephalic astrocytes reveals adult astrogenesis regulated by Smad4 (2021)
Figure 8

Figure 8 - RNA-seq of astrocytes from adult mouse cerebral cortex grey matter and diencephalon

A Schematic drawing of Ribotag assisted mRNA isolation from astrocytes isolated from regions dissected as indicated in red (CTX GM) and blue (DIE).

B Micrographs of forebrain sections of 3 mo old GLASTCreERT2/RPL22HA/eGFP 21 dpt mice stained for HA show maximum intensity projections of HA+ expressing cells with astroglial morphology. CTX GM: cerebral cortex grey matter; DIE: diencephalon. Scale bars: 100 µm.

C Micrographs of forebrain sections of 3 mo old GLASTCreERT2/RPL22HA/eGFP mice 21dpt stained for HA and S100β showing the specificity of the mouse line with HA detected only in S100β+ astrocytes. DIE: diencephalon. Scale bar: 70 μm.

D Histogram showing expression levels of the top 10 astrocyte specific markers genes in DIE (blue bars) and CTX GM (red bars) samples.

E Principal Component Analysis (PCA) from DeSeq2 expression data shows distinct clusters of samples from 3 DIE (blue) and 3 CTX GM (red) astrocyte sample preparations.

F Venn diagram depicting co-expressed and differentially expressed gene numbers in DIE and CTX GM astrocytes (see Dataset EV4 for Figure 8).

G, H Heatmaps depicting top 40 differentially expressed genes (G, row scaled log normalized read counts) or transcription factors (H, Library size normalized log2 transformed read counts; from DIE and CTX GM astrocytes. Transcription factors were identified according to the content of AnimalTFDB database (http://www.bioguo.org/AnimalTFDB/).

I Bar chart depicting significantly enriched GO terms (analyzed first by Gorilla followed by Revigo) based on the 379 differentially expressed transcription factor genes (DeSeq2, cutoff pAdj0.01) comparing DIE and CTX GM astrocytes. Terms related to cell division, cell proliferation etc. are highlighted in red. (P-value threshold was set to 0.01 http://cbl-gorilla.cs.technion.ac.il).

Data information: CTX GM: cerebral cortex grey matter; DIE: diencephalon; n=3 biological replicates, tissue from 3 mice was pooled
Molecular diversity of diencephalic astrocytes reveals adult astrogenesis regulated by Smad4 (2021)
Figure 9

Figure 9 - Smad4 regulates diencephalic astrocyte proliferation and neurosphere formation

A Reactome Pathway analysis for all 174 transcription factor genes with higher expression in DIE (upper) and all 205 genes higher in CTX GM (lower) astrocytes.

B t-SNE visualization of DIE astrocytes isolated by ACSA-2 MACS (Figure 1A) showing Smad4 expression (scaled log normalized read counts).

C Example of a Smad4/GFP double-positive astrocyte (indicated by arrowhead) in a single optical section of DIE from 3 mo old GLASTCreERT2/eGFP mice 21 dpt immunostained as indicated on top of the panel. Orthogonal projections of the area highlighted in left panel of C shown at higher magnification in the right panel. Scale bar: 15 µm.

D Example of a proliferating (PCNA+, arrowhead) astrocyte in a single optical section of DIE from 3 mo old GLASTCreERT2/eGFP mice 21 dpt immunostained as indicated on top with single channels shown in middle and right panels. Scale bars: 15 µm.

E Histogram depicting percentage of PCNA positive cells of all GFP positive cells in the DIE of the genotype indicated on the x-axis (n = 6 animals per genotype; 5 ROI covering the diencephalon on a total of 3 slides were analyzed per animal; PCNA negative and positive cells comparing wt/wt vs. fl/fl animals, adjusted P-value = 0.002).

F, G Histograms depicting percentage of the neurosphere-generating cells obtained from the DIE (F) or SEZ (G) of GLASTCreERT2/Smad4fl/fl/eGFP or GLASTCreERT2/Smad4WT/WT/eGFP, as indicated on the x-axis (n = 3-4 animals for SEZ, 4 to 8 technical replicates analyzed per animal; 4 animals for DIE, 8 technical replicates analyzed per animal).

H Histogram depicting the number of secondary neurospheres generated from 10000 Smad4f/f neurosphere cells at 7 days after infection with MLV-based retrovirus containing either CAG-NLS-Cre or only CAG-GFP (n = 11 cultures / animals, 3 technical replicates per condition analyzed; control vs. Cre, P-value = 0.0149). Note the decrease in DIE astrocyte proliferation in vivo (E) and in neurosphere formation upon Smad4 deletion (F, H).

Data information: In E, F, G and H data are presented as mean ± SEM. F: GFP negative and GFP positive cells comparing wt/wt vs fl/fl animals, adjusted P-value = 0.0408; G: GFP negative and GFP positive cells comparing wt/wt vs fl/fl animals, ns). Each dot represents one n. * P < 0.05, ** P < 0.01 (E, F, G: nonparametric 2way ANOVA followed by Sidak's multiple comparisons test, H: nonparametric Mann-Whitney test).

Expanded View Figure Legends
Legionella- and host-driven lipid flux at LCV-ER membrane contact sites promotes vacuole remodeling (2022)
Figure 1

Figure 1. Dictyostelium discoideum MCS components localize to LCVs and/or ER.

(A) Dually labeled D. discoideum amoebae producing MCS components fused to GFP, and either calnexin (CnxA)-mCherry (pAW012), P4C-mCherry (pWS032), or AmtA-mCherry were infected (MOI 5, 2 h) with mCerulean-producing L. pneumophila JR32 (pNP99), fixed with 4 % PFA, and imaged by confocal fluorescence microscopy. Merged images are shown. Scale bars: 3 μm (insets: 1 μm).

(B-D) The Pearson's correlation coefficients were generated using Coloc 2 from Fiji (ImageJ) and are shown for MCS components fused to GFP with respect to (B) CnxA-mCherry (n=146), (C) P4C-mCherry (n=138), or (D) AmtA-mCherry (n=159). Data represent mean and SEM of three independent biological replicates (statistics refer to JR32-infected D. discoideum producing GFP-Sac1; *P<0.05; **P<0.01; ***P<0.001, Student`s t-test).
Legionella- and host-driven lipid flux at LCV-ER membrane contact sites promotes vacuole remodeling (2022)
Figure 2

Figure 2. Vap colocalizes with OSBP8 and OSBP11 at the LCV-ER MCS.

(A) Dually labeled D. discoideum Ax3 producing mCherry-Vap (pSV048) and OSBP8-GFP (pMIB89) or GFP-OSBP11 (pMIB39) were infected (MOI 5, 1-8 h) with mCerulean-producing L. pneumophila JR32 (pNP99) and fixed with 4 % PFA. Merged images for the analyzed time points are shown. Scale bars: 3 μm (insets: 1 μm).

(B) The Pearson's correlation coefficient was generated using Coloc 2 from Fiji (ImageJ) and is shown for OSBP8 or OSBP11 fused to GFP with respect to mCherry-Vap (n=100-150 each per time point). Data represent mean and SEM of three independent biological replicates (**P<0.01, Student`s t-test).
Legionella- and host-driven lipid flux at LCV-ER membrane contact sites promotes vacuole remodeling (2022)
Figure 4

Figure 4. Vap, OSBP11, and Sac1 promote expansion of PtdIns(4)P-positive LCVs.

(A) Dually labeled D. discoideum Ax3, ∆vap, ∆osbG, ∆osbH, ∆osbK or ∆sey1 mutants producing P4C-mCherry (pWS032) and CnxA-GFP (pAW016), or Ax3 producing P4C-mCherry and either GFP-Sac1 (pLS037), GFP-Sac1_C383S (pSV015) or GFP-Sac1_ΔTMD (pSV034) were infected (MOI 5, 2-16 h) with mCerulean-producing L. pneumophila JR32 (pNP99) and fixed with 4 % PFA. Merged images for the analyzed time points are shown. Scale bars: 3 μm.

(B) LCV area was measured using ImageJ (n=100-200 per condition from 3 independent biological replicates). Means and SEM of single cells are shown (**P<0.01; ***P<0.001, Student`s t-test).
Legionella- and host-driven lipid flux at LCV-ER membrane contact sites promotes vacuole remodeling (2022)
Figure 5

Figure 5. Sac1_ΔTMD reduces the accumulation of PtdIns(4)P, Vap and OSBP8 on LCVs.

(A) Dually labeled D. discoideum Ax3, ∆vap, ∆osbG, ∆osbH, ∆osbK or ∆sey1 mutants producing P4C-mCherry (pWS032) and GFP-Sac1 (pLS037) were infected (MOI 5, 2-16 h) with mCerulean-producing L. pneumophila JR32 (pNP99) and fixed with 4 % PFA. Merged images for the analyzed time points are shown. Scale bars: 3 μm.

(B, C) Imaging flow cytometry (IFC) analysis of dually labeled D. discoideum Ax3, ∆vap, ∆osbG, ∆osbH, ∆osbK or ∆sey1 mutants producing P4C-mCherry (pWS032) and either GFP-Sac1 (pLS037) or GFP-Sac1_ΔTMD (pSV034), infected (MOI 5, 2 h) with mPlum-producing L. pneumophila JR32 (pAW014).

(D) Dually labeled D. discoideum Ax3 producing GFP fusions of MCS components and either mCherry-Sac1 (pSV044) or mCherry-Sac1_ΔTMD (pSV045) were infected (MOI 5, 2 h) with mCerulean-producing L. pneumophila JR32 (pNP99) and fixed with 4 % PFA. Merged images for the analyzed time points are shown. Scale bars: 3 μm.

(E) IFC analysis of dually labeled D. discoideum Ax3 producing GFP fusions of MCS components and either mCherry-Sac1 (pSV044) or mCherry-Sac1_ΔTMD (pSV045), infected (MOI 5, 2 h) with mPlum-producing L. pneumophila JR32 (pAW014). Quantification of (B) P4C-mCherry (C) Sac1-GFP or (E) GFP fusions of MCS components localizing to LCVs at 2 h p.i. Due to the lower resolution of IFC, "LCVs" designates the LCV limiting membrane and tightly attached ER. Number of events per sample, n = 5000.

Data information (B, C, E): Data represent mean and SEM of three independent biological replicates (*P<0.05; **P<0.01; ***P<0.001, Student`s t-test).
Legionella- and host-driven lipid flux at LCV-ER membrane contact sites promotes vacuole remodeling (2022)
Figure 6

Figure 6. PtdIns(4)P and sterol exchange at LCV-ER MCS.

(A) Dually labeled D. discoideum Ax3 producing GFP-D4H* (pSV046) or P4C-GFP (pWS034) and P4C-mCherry (pWS032), CnxA-mCherry (pAW012), or AmtA-mCherry were infected (MOI 5, 15 - 120 min) with mCerulean-producing L. pneumophila JR32 (pNP99) and fixed with 4 % PFA. Merged images for the analyzed time points are shown. Scale bars: 3 μm.

(B-C) The Pearson's correlation coefficient was generated using Coloc 2 from Fiji (ImageJ) and is shown for (B) AmtA-mCherry with respect to P4C-GFP or GFP-D4H* (n=100-150 per time point), and (C) GFP-D4H* with respect to P4C-mCherry or CnxA-mCherry (n=100-150 each per time point). Data represent mean and SEM of three independent biological replicates (*P<0.05, Student`s t-test).

(D) Single labeled D. discoideum Ax3 producing either CnxA-GFP (pAW016) or P4C-GFP (pWS034) were infected (MOI 5, 15-120 min) with mCherry-producing L. pneumophila JR32 (pNP102), fixed with 4 % PFA and stained with filipin. Merged images for the analyzed time points are shown. Scale bars: 3 μm.

(E) The Pearson's correlation coefficient was generated using Coloc 2 from Fiji (ImageJ) and is shown for filipin with respect to CnxA-GFP or P4C-GFP (n=80-90 each per time point). Data represent mean and SEM of the means of three independent biological replicates (***P<0.001, Student`s t-test).
Legionella- and host-driven lipid flux at LCV-ER membrane contact sites promotes vacuole remodeling (2022)
Figure 7

Figure 7. The L. pneumophila effectors LepB and SidC determine PtdIns(4)P enrichment and expansion of LCVs.

(A) Dually labeled D. discoideum Ax3 producing P4C-mCherry (pWS032) and either CnxA-GFP (pAW016), GFP-Sac1 (pLS037), or GFP-Sac1_ΔTMD (pSV034) were infected (MOI 5, 1-8 h) with mCerulean-producing L. pneumophila JR32, ΔlepB or ΔsidC (pNP99) and fixed with 4 % PFA. Merged images for the analyzed time points are shown. Scale bars: 3 μm.

(B) LCV area was measured using ImageJ (n=100-200 per condition). Means and SEM of single cells are shown (**P<0.01; ***P<0.001, Student`s t-test).

(C) Imaging flow cytometry (IFC) analysis of dually labeled D. discoideum Ax3 producing P4C-mCherry (pWS032) and either CnxA-GFP (pAW016), GFP-Sac1 (pLS037), or GFP-Sac1_ΔTMD (pSV034), infected (MOI 5, 1-8 h) with mPlum-producing L. pneumophila JR32 (pAW014). Number of events per sample, n = 5000. Data represent mean and SEM of three independent biological replicates (*P<0.05; **P<0.01; ***P<0.001, Student`s t-test).
The CD38-NADase is a new major contributor to Duchenne muscular dystrophic phenotype (2022)
Figure 1

Figure 1. Restoration of NAD+ levels in mdx/CD38-/- mice

A,B NAD and nicotinamide (NAM) levels in heart of WT (n=5, n=6, respectively), mdx (n=5, n=5, respectively) and mdx/CD38-/- mice (n=6, n=5, respectively).

C,D NAD and NAM levels in diaphragm of WT (n=5, n=6, respectively), mdx (n=4, n=5, respectively) and mdx/CD38-/- mice (n=6, n=5, respectively).

E,F,G levels of ADP-ribose (ADPR) (E), cyclic ADP-ribose (cADPR) (F) expressed as nmol/mg protein and (G) qPCR analysis of mRNA levels of Sterile alpha and Toll/interleukin-1 receptor motif-containing 1 (SARM1) in heart of WT (n=6), mdx (n=5) and mdx/CD38-/- mice (n=5, n=5, n=6 respectively).

H,I,J Levels of ADPR (H), cADPR (I) expressed as nmol/mg protein and (J) qPCR analysis mRNA levels of SARM1 in diaphragm of WT (n=6), mdx (n=5) and mdx/CD38-/- (n=5) mice.

K, L Western blot analysis of CD38 protein expression in heart (K) and diaphragm (L) of WT (n=5, n=6, respectively) and mdx (n=5) mice. Vinculin is used as housekeeping protein control and the dot plots show the ratio of CD38/vinculin.

M, N qPCR analysis of CD38 mRNA levels in heart (M) and diaphragm (N) of WT (n=6) and mdx (n=5) mice.

Data information: A-N Each dot of the graphs represents a mouse and measured in duplicate except for K,L, a value/mouse. After normality and variance comparison tests, significance was assessed using: A,C,E,F,G,H: ANOVA followed by Fisher's LSD test; B: Kruskal-Wallis followed by Dunn's test; D: Kruskal-Wallis followed by Mann-Whitney tests; I: Welch's ANOVA followed by Welch's t-tests; J: ANOVA; K,L,M: unpaired Student's t-test; N: unpaired Welch's t-test. Values are expressed as means ± SEM. Significance: *p<0.05, **p<0.01, ***p<0.001.
The CD38-NADase is a new major contributor to Duchenne muscular dystrophic phenotype (2022)
Figure 2

Figure 2. Deletion of CD38 prevented heart dysfunction and reduces spontaneous Ca2+ activity in isolated cardiomyocytes of mdx/CD38-/- mice

A Cardiac function evaluated by echocardiography in mdx/CD38-/- mice: the dot plots show the left ventricular (LV) diastolic (A1) and systolic (A2) inner diameters and LV ejection (A3) fraction in 7-month-old WT (n=5), mdx (n=6) and mdx/CD38-/- (n=10) mice.

B Plasma levels of cardiac stress biomarkers: brain natriuretic peptide (BNP) (B1) and cardiac troponin I (cTnI) (B2) in 7-month-old WT (n=7), mdx (n=6) and mdx/CD38-/- (n=11) mice.

C Cropped images revealed the collagen (blue) stained by Masson's trichrome staining in the heart of WT, mdx, and mdx/CD38-/- mice. Dot plot showing the quantification of heart collagen staining area (% total area) in 7-month-old WT, mdx, and mdx/CD38-/- mice (n=4 per group). Scale bars: 200 µm.

D Isoproterenol-induced heart failure in 3-month-old mice. The Kaplan-Meier curve shows the survival rate of mdx mice (NaCl, n=5); mdx (n=19) and mdx/CD38-/- (n=9) mice following isoproterenol (subcutaneous injection) at 2.5 mg/kg/d for 10 days.

E Histogram showing isoproterenol-induced heart hypertrophy in mdx NaCl (n=5) mice, surviving mdx (n=9), mdx/CD38-/- NaCl (n=9) and mdx/CD38-/- (n=9) mice, expressed as heart weight/body weight ratio (HW/BW).

F Plasma levels of cardiac stress biomarkers: brain natriuretic peptide (BNP) (F1) and cardiac troponin I (cTnI) (F2) following isoproterenol treatment in mdx NaCl (n=5), surviving mdx (n=8), mdx/CD38-/- NaCl (n=5) and mdx/CD38-/- (n=8) mice.

G,H Time-lapse images recorded by confocal microscopy in "line scanning mode", showing Ca2+ sparks and waves in cardiomyocytes at rest, extracted from mdx and mdx/CD38-/- mice; scale bars: 10 µM (horizontal), 500 ms (vertical). Bar graphs showing the averaged Ca2+ sparks (G) and waves (H) frequencies in cardiomyocytes isolated from WT (n=21 cells), mdx (n=28 cells) and mdx/CD38-/- (n=32 cells) mice.

I Bar graph showing the fractional release following caffeine application in cardiomyocytes from WT (n=24 cells), mdx (n=33 cells) and mdx/CD38-/- (n=32 cells) mice.

J Bar graph showing the post-rest potentiation in cardiomyocytes from WT (n=24 cells), mdx (n=28 cells) and mdx/CD38-/- (n=32 cells) mice.

Data information: A,B,C: Each dot of the graphs represents a mouse. A,F in duplicate; C in triplicate; D,E one value/mouse. G,H,I,J: experiments performed in 3 mice of each group. After normality and variance comparison tests, significance was assessed using:A1,B1,B2,C,I: ANOVA followed by Fisher's LSD test; A2,A3,G,H,J: Kruskal-Wallis followed by Dunn's test; D: Log-rank Mantel-Cox test and Log-rank test for trend and Gehan-Breslow-Wilcoxon test; E,F2 Kruskal-Wallis followed by Mann-Whitney tests; F1: ANOVA followed by Student's/ Welch's t-tests; Values are expressed as means ± SEM. Significance: *p<0.05, **p<0.01, ***p<0.001.
The CD38-NADase is a new major contributor to Duchenne muscular dystrophic phenotype (2022)
Figure 3

Figure 3. Deletion of CD38 improved diaphragm structure and function in mdx/CD38-/- mice

A Measurement of the ventilatory mechanic by barometric plethysmography: dot plots showing inspiratory (A1) and expiratory times (A2), the relaxation time (A3) and the respiratory frequency (A4) in WT (n=8), mdx (n=11) and mdx/CD38-/- (n=9) mice.

B Images showing muscle fiber typology revealed by immunostaining. Localization of the slow MyHC (Type I) fiber and fast MyHCs (Type IIa and IIb/X) fibers, along with laminin (red) on transverse cross-sections from diaphragm of WT, mdx and mdx/CD38-/- mice. Scale bars: 100 µm. Histogram showing the percentage of I, IIa and IIb/X fiber type distribution in diaphragm of WT (n=7), mdx (n=6) and mdx/CD38-/- (n=7) mice.

C Fiber size distribution in the diaphragm of WT (n=7), mdx (n=6) and mdx/CD38-/- (n=7) mice.

D Images revealing the collagen (blue) by Masson's trichrome staining in diaphragm of WT, mdx, and mdx/CD38-/- mice. Quantification of collagen staining area (% total area) in the diaphragm of WT, mdx and mdx/CD38-/- mice (n=4 per group). Scale bars: 200 µm.

E Embryonic myosin expression: immunostaining showing its localization along with laminin (green) on transverse cross-sections from diaphragm of WT, mdx and mdx/CD38-/- mice. Scale bars: 50 µm. Histogram showing the relative proportion of embryonic myosin area (% total area) in diaphragm of WT (n=7), mdx (n=8) and mdx/CD38-/- (n=8) mice.

Data information: Each dot of the graphs (A,D,E) represents a mouse. A,B,C one value/mouse; D in duplicate; E one value/mouse. After normality and variance comparison tests, significance was assessed using: A1,A3,D: ANOVA followed by Fisher's LSD test; A2, A4 : Kruskal-Wallis followed by Dunn's test; B: Chi-square test C: Kolmogorov-Smirnov test; E: Welch's ANOVA followed by Welch's t-tests. Values are expressed as means ± SEM. Significance: *p<0.05, **p<0.01, ***p<0.001. WT images in 3B and 3E have been redisplayed in S4B and S4E, respectively.
The CD38-NADase is a new major contributor to Duchenne muscular dystrophic phenotype (2022)
Figure 4

Figure 4. Deletion of CD38 reduced cell infiltrations and inflammation markers in diaphragm in mdx/CD38-/- mice

A Images showing immunostaining of myeloid cells showing F4/80 (macrophage marker), Ly-6G/6C (monocyte, granulocyte and neutrophil marker), CD8 (cytotoxic T lymphocyte marker) and IL-6 positive cells in diaphragm of 7-month-old WT (n=8), mdx (n=8) and mdx/CD38-/- (n=7 or 8 ) mice. Lower panel: histograms quantifying the percentage of F4/80 (A1) mdx/CD38-/- (n=7), LY6 (A2) mdx/CD38-/- (n=8), CD8 (A3) mdx/CD38-/- (n=7), IL-6 (A4) mdx/CD38-/- (n=8) positive cells. Scale bars: 50 µm.

B qPCR analysis of mRNA levels of interleukin-1β (IL-1ß) (B1) and -6 (IL-6) (B2), cyclin-dependent kinase inhibitor 1 (p21) (B3) transforming growth factor-β (TGF-β) (B4), and senescence markers (cell-cycle inhibitor p16, INK4a) (B5), Collagen Type I Alpha 1 Chain (Col1a1) (B6) in diaphragm of 20-month-old WT (n=6), mdx (n=5) and mdx/CD38-/- (n=5) mice.

Data information: A,B Each dot of the graphs represents a mouse. A one value/mouse; B in duplicate. After normality and variance comparison tests, significance was assessed using: A1-4: Welch's ANOVA followed by Welch's t-tests; B1-6: ANOVA followed by Fisher's LSD test. Values are expressed as means ± SEM. Significance: *p<0.05, **p<0.01, ***p<0.001.
The CD38-NADase is a new major contributor to Duchenne muscular dystrophic phenotype (2022)
Figure 5

Figure 5. Deletion of CD38 improved skeletal muscle structure and function in mdx/CD38-/- mice

A NAD+ levels in limb muscles of 20-month-old WT (n=7), mdx (n=5) and mdx/CD38-/- (n=6) mice.

B CD38 expression: western blot analysis of CD38 (B1) protein in limb of WT and mdx mice (n=6 per group). Vinculin is used as housekeeping protein control and the dot plots show the ratio of CD38/vinculin. qPCR analysis of CD38 mRNA (B2) in limb of 20-month-old WT and mdx mice (n=4 per group).

C Histograms showing the grip duration (latency to fall) (C1) and the limb force (C2) measured by a grip test in WT (n=89), mdx (n=52, 53 respectively) and mdx/CD38-/- (n=58) mice (age: 9 to 26 months old).

D Images showing muscle fiber typology revealed by immunostaining. Localization of the slow MyHC (Type I) fiber and fast MyHCs (Type IIa and IIb/X) fibers, along with laminin (red) on transverse cross-sections from soleus and tibialis (TA) of WT, mdx and mdx/CD38-/- mice. Scale bars: 100 µm. Histogram showing the percentage of I, IIa and IIb/X fiber type distribution in soleus and TA of WT, mdx and mdx/CD38-/- mice. Experiments performed in WT (n=3), mdx (n=5) and mdx/CD38-/- (n=7) mice for soleus; WT (n=5), mdx (n=6) and mdx/CD38-/- (n=6) mice for TA.

E Fiber size distribution in soleus from WT, mdx and mdx/CD38-/- mice.

F Images displaying the collagen (blue) revealed by Masson's trichrome staining in limb of WT, mdx, and mdx/CD38-/- mice. The dot plot shows the quantification of collagen staining area (% total area) in limb of WT (n=4), mdx (n=5) and mdx/CD38-/- (n=5) mice. Scale bars: 200 µm.

G Embryonic myosin expression revealed by immunostaining along with laminin (green) on transverse cross-sections from limb of WT (n=5), mdx (n=8) and mdx/CD38-/- (n=8) mice. Scale bars: 50 µm. Dot plot showing the relative proportion of embryonic myosin area in limb of WT, mdx and mdx/CD38-/- mice.

Data information: Each dot of the graphs represents a mouse. B,C1,D,E,G one value/mouse; A in duplicate; F,C2 in triplicate. After normality and variance comparison tests, significance was assessed using: A,F: ANOVA followed by Fisher's LSD test; B1: unpaired Welch's t-test; B2 : unpaired Student's t-test; C1-2 : Kruskal-Wallis followed by Dunn's test; D: Chi-square test; E: Kolmogorov-Smirnov test; G: Welch's ANOVA followed by Welch's t-tests. Values are expressed as means ± SEM. Significance: *p<0.05, **p<0.01, ***p<0.001. WT images in 5D and 5G have been redisplayed in S6B and S6E, respectively.