imaris software analysis colocalization  (Oxford Instruments)

Journal: bioRxiv

Article Title: Loss of Parkin Disrupts Nuclear and Mitochondrial Programs Required for Muscle Regeneration

doi: 10.64898/2026.03.20.712989

Figure Lengend Snippet: A-B ) Volume of mitochondria colocalized to autophagosomes expressed in absolute value or relative to total mitochondrial volume in in situ fixed, freshly sorted quiescent and 4h in vitro activated MuSCs ( n =48-55 cells from 3 mice in each group and experimental condition). C) Confocal image and 3D reconstruction of TOM20-labeled mitochondria (green) and LC3-labeled autophagosomes (red) in each of the experimental conditions examined. The volume of mitochondria overlapping with autophagosomes is shown in yellow in the right-end panels, where mitochondria and autophagosomes surfaces have been removed to highlight changes in colocalization. D) Colocalization of PARKIN to TOM20-labeled mitochondria expressed in absolute volume of PARKIN + structures overlapping with mitochondria per cell ( n =25-36 cells from 3 mice in each group and experimental condition). E) Proportion of TOM20-labeled mitochondria labeled with PARKIN, computed using data presented in panel D and total mitochondrial content (Fig. S1A). F) Confocal image and 3D reconstruction of TOM20-labeled mitochondria (green) and PARKIN-labeled structures (red) in each of the experimental conditions examined. The volume of PARKIN overlapping with mitochondria is shown in yellow in the right-end panels, where mitochondria and PARKIN + surfaces have been removed to highlight changes in colocalization. G) Total cellular PARKIN content in the indicated experimental conditions. H) Parkin transcript levels in MuSCs purified from muscles that were fixed in situ in healthy uninjured conditions or at 15, 30, 60, 90 and 120 min after CTX injury. Data is taken from (GSE163856 ). All data are presented as mean ± SEM. ns: not significant, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0001 on unpaired two-tailed t tests or one-way ANOVAs. I) Changes in the transcript levels of ubiquitin- and receptor-dependent mitophagy in three transcriptomics datasets (GSE70736, GSE55490, GSE47177 , , ) that compared quiescent and in vivo activated MuSCs 36-72h following muscle injury with CTX or BaCl 2 . Each dot represents data from an individual dataset and a specific timepoints. Genes shown where differentially expressed in all datasets with a q value of less than 0.05. Level of statistical significance shown is based on the average q value.

Article Snippet: Volume reconstruction and surface/surface colocalization analysis was performed in Imaris using fixed settings.

Techniques: In Situ, In Vitro, Labeling, Purification, Muscles, Two Tailed Test, Ubiquitin Proteomics, Transcriptomics, In Vivo

Journal: bioRxiv

Article Title: Loss of Parkin Disrupts Nuclear and Mitochondrial Programs Required for Muscle Regeneration

doi: 10.64898/2026.03.20.712989

Figure Lengend Snippet: A-C) Parkin transcript and protein levels in FACS-purified MuSCs from MuSC Park2 +/+ and MuSC Park2 -/- mice 1 week after the last tamoxifen treatment ( n =4 mice per group). Protein abundance was quantified as the total volume of PARKIN + structures (B) in cells labeled with antibodies against PARKIN and TOM20 ( n =11-70 cells from 2-4 mice per group,) (C). D-E) transcript abundance expressed in Transcript Per Million (TPM) for genes involved in Parkin- and Receptor-mediated mitophagy ( n =3 mice per group). F-G) Volume of mitochondria colocalized to autophagosomes expressed in absolute value or relative to total mitochondrial volume in freshly sorted quiescent and 4h in vitro activated MuSCs ( n =41-52 cells from 3 mice in each group). H) Confocal image and 3D reconstruction of TOM20-labeled mitochondria (green) and LC3-labeled autophagosomes (red) in each of the experimental conditions examined. The volume of mitochondria overlapping with autophagosomes is shown in yellow in the right-end panels, where mitochondria and autophagosomes surfaces have been removed to highlight changes in colocalization. All data are presented as mean ±SEM. ns: not significant, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0001 on unpaired two-tailed t tests or one-way ANOVAs.

Article Snippet: Volume reconstruction and surface/surface colocalization analysis was performed in Imaris using fixed settings.

Techniques: Purification, Quantitative Proteomics, Labeling, In Vitro, Two Tailed Test

Journal: Nature Neuroscience

Article Title: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models

doi: 10.1038/s41593-025-02062-6

Figure Lengend Snippet: a ) Representative immunofluorescent images of TDP-43 localization in NMJs from gastrocnemius muscle of induced TDP43∆NLS (-Dox; 2-weeks) and non-induced control (+Dox). Gray indicates TDP-43 and TDP-43:NFH 3D colocalization result (in the respective panel), blue indicates NFH, red indicates BTX. Scale bar: 10 µm. experiment repeated in 5 mice per genotype. b ) Representative immunofluorescent images of pTDP-43 localization in NMJs from EDL muscle of p60 B6SJL.SOD1 G93A and littermate control mice. Gray indicates pTDP-43 and pTDP-43:NFH 3D colocalization result (in the respective panel), blue indicates NFH, red indicates BTX. Scale bar: 10 µm. experiment repeated in 3 mice per genotype. c ) Representative immunofluorescent images of pTDP-43 localization in NMJs from EDL muscle of p290 SOD1 G37R and littermate control mice. Gray indicates pTDP-43 and pTDP-43:NFH 3D colocalization result (in the respective panel), blue indicates NFH, red indicates BTX. Scale bar: 10 µm. experiment repeated in 3 mice per genotype. d, e ) Representative immunofluorescent images ( d ) and quantitative analysis ( e ) of TDP-43 in NMJs from EDL muscle of p290 SOD1 G37R and littermate control mice. Gray indicates TDP-43 and TDP-43:NFH 3D colocalization result (in the respective panel), blue indicates NFH, red indicates BTX. Scale bar: 10 µm. Data are shown in violin density plots with markings of first, median and third quartiles of mean TDP-43 intensity to NFH/Synaptophysin intensity in NMJs. Two-tail unpaired student’s t-test. ****p = 2.57×10 −5 . n = 38 NMJs from 3 mice per condition. f ) Quantitative analysis of the percent of pTDP-43 colocalization area in presynapses at NMJs of P60 B6SJL.SOD1 G93A . Two-tail unpaired student’s t-test. **p = 0.0071. n = 33;21 NMJs from 3 mice per condition. g, h ) Representative immunofluorescent images ( g ) and quantitative analysis ( h ) of pTDP-43 in NMJs from EDL muscle of p90 B6SJL.SOD1 G93A and littermate control mice. Gray indicates pTDP-43 and pTDP-43:NFH 3D colocalization result (in the respective panel), blue indicates NFH, red indicates BTX. Scale bar: 10 µm. Data are shown in violin density plots with markings of first, median and third quartiles of mean pTDP-43 intensity to NFH intensity (left panel) or percent of pTDP-43 colocalization area in presynapses at NMJs. Two-tail unpaired student’s t-test. **p = 0.0021(pTDP-43 intensity), ****p = 4.8×10 −9 (pTDP-43 area). n = 39;36 NMJs from 3 mice per condition. i, j ) Representative immunofluorescent images of TDP-43 ( i ) and pTDP-43 ( j ) localization in NMJs from soleus muscles of p290 SOD1 G37R and littermate control mice. Gray indicates TDP-43 and TDP-43:NFH 3D colocalization result (in the respective panel), blue indicates NFH, red indicates BTX. Scale bar: 10 µm. k ) Quantitative analysis for the percent of NMJs with apparent TDP-43/pTDP-43 patch in soleus muscles from p290 SOD1 G37R and littermate controls. Data are shown as the mean percent of NMJs with TDP-43 (left panel) or pTDP-43 (right panel) patch ± SEM. n = 3 mice per group. Two-tailed, unpaired t-test. p = 0.0746(TDP-43), p = 0.0636(pTDP-43). l ) Quantitative analysis of TDP-43 (left panel) and pTDP-43 (right panel) intensity in NMJs from soleus muscle of p290 SOD1 G37R and littermate control mice. Data are shown in violin density plots with markings of first, median and third quartiles of mean TDP-43 (left) or pTDP-43 (right) intensity to NFH/Synaptophysin intensity in NMJs. Two-tail unpaired student’s t-test. n = 44;41 (TDP-43), n = 82,52 (pTDP-43) NMJs from 3 mice per condition. ****p = 5.295×10 −5 (TDP-43), p = 0.0899(pTDP-43).

Article Snippet: ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle.

Techniques: Control, Muscles, Two Tailed Test

Journal: Nature Neuroscience

Article Title: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models

doi: 10.1038/s41593-025-02062-6

Figure Lengend Snippet: a ) Relative mRNA transcript levels in axons versus cell-bodies (soma) of Tardbp , Elavl1 , Stau1 , and Actb . Data are shown as fold change (F.C) of normalized counts in axons over soma for each mRNA transcript. Raw data reanalysis of axonal and soma RNA-seq from primary motor neurons (Rotem et al., 2017). b ) Representative images of smFISH for β-actin and TDP-43 mRNAs compared with no probe control in primary motor neuron cell bodies. Gray indicates RNA FISH, green indicates phalloidin/HB9:GFP, blue indicates nuclei (DAPI). Red dashed line marks proximal axon shown in magnified insets. Scale bar: 15 µm. Experiment performed in 3 neuronal cultures. c-d ) Representative images ( c ) and quantitative analysis ( d ) of TDP-43 immunolabeling of distal axons in MFC 4 days after localized transfection with TDP-43 siRNA mix or with control siRNA. Magenta indicates TDP-43, green indicates NFH. Scale bar: 10 µm. Data are shown in violin density plots with markings of first, median and third quartiles of mean number of TDP-43 puncta in 100 µm axons. Two-tailed unpaired student’s t-test. ****p < 1 × 10 −14 . n = 94;87 axons from 3 independently grown co-cultures. e ) Representative immunofluorescent images of TDP-43 puro-PLA in a growth cone at a premature NMJ, and in a mature NMJ in healthy co-cultures. Blue indicates NFH, gray indicates BTX, yellow indicates TDP-43 puro-PLA, white dashed line indicates skeletal muscle margins (in NMJ panel, the entire region is over a skeletal muscle) Scale bar: 5 µm. Experiment repeated in 3 neuromuscular co-cultures.

Article Snippet: ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle.

Techniques: RNA Sequencing, Control, Immunolabeling, Transfection, Two Tailed Test

Journal: Nature Neuroscience

Article Title: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models

doi: 10.1038/s41593-025-02062-6

Figure Lengend Snippet: a – c , Representative images and quantitative analysis ( b , c ) of NMJ immunolabeling for CD63 (upper panel b ) and CHMP2A (lower panel c ). Gray indicates three-dimensional co-localization result of CD63/CHMP2A and BTX. Scale bar, 10 µm. n = 19 ( b ) and n = 39 ( c ) muscles. *** P = 0.00012 ( b ), **** P = 1.17 × 10 −10 ( c ). d , Illustration of experimental setup in e and f . Primary skeletal muscles were transfected with CD63-pHluorin vector and co-cultured with primary MNs in compartmental MFCs. e , f , Representative images and quantification of CD63-pHluorin signal and localization in neuromuscular co-cultures. Scale bar, 20 µm. n = 24 muscles from three independent repeats. Two-tailed paired Student’s t -test, ** P = 0.0015. g , Representative TEM images of muscle-derived EVs. Scale bars, 300 nm (left panel) and 100 nm (right panel). h , Representative NTA plot for muscle conditioned media. i , Representative images of western blots for CD63 (55 kD), CD81 (26 kD), CD9 (22 kD), Ago2 (87 kD) and TDP-43 (43 kD) in protein lysates of primary muscles and primary muscle-derived EVs. j , Illustration of experimental setup for j . Primary skeletal muscles were transfected with Ago2–GFP vector and co-cultured with primary MNs in compartmental MFCs. k , Upper panel: representative images of immunolabeling for Ago2 and GFP in neuromuscular co-cultures. Lower panel: inset and representative image of Imaris puncta analysis for Ago2 and GFP antibody labeling within axons. Yellow arrowheads indicate co-localized GFP and Ago2 signals in axons. Scale bar, 10 µm. For b , c , f , data are shown as pairs of signal intensity in synaptic versus extra-synaptic regions within the same muscle. For b , c , experiment was repeated in three mice. For f , experiment was repeated in three co-cultures. For b , c , f , two-tailed paired Student’s t -test. For g , h , I , representative for three biological repeats of muscle culture and EV preparation. For k , repeated once. Representative of 15 images. coloc, co-localization; Ex.synaptic, extra-synaptic.

Article Snippet: ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle.

Techniques: Immunolabeling, Muscles, Transfection, Plasmid Preparation, Cell Culture, Two Tailed Test, Derivative Assay, Western Blot, Antibody Labeling

Journal: Nature Neuroscience

Article Title: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models

doi: 10.1038/s41593-025-02062-6

Figure Lengend Snippet: a ) Representative images for the localization of CD63 (upper panel), CD81 (mid panel) and CHMP2A (lower panel) in healthy GC muscle NMJs. Blue indicates NFH, red indicates BTX, Dynamic blue-green-yellow indicates CD63/CD81/CHMP2A according to raw fluorescent intensity (blue=low; yellow=high). Scale bar: 10 µm. Experiment repeated 3 times (CD63, CD81) and 6 times for CHMP2A. b ) Representative images for the localization of CD63 (upper panel), CD81 (lower panel) in NMJs within dissociated muscle fibers following enzymatic digestion of surrounding cells and connective tissues. Red indicates BTX, Dynamic blue-green-yellow indicates CD63/CD81 according to raw fluorescent intensity (blue=low; yellow=high). Scale bar: 10 µm. Experiment repeated 3 times. c ) Quantitative analysis of CD81 raw fluorescent signal in synaptic versus extrasynaptic regions within the same muscle fiber. Two-tail paired student’s t-test. ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle. White indicates BTX, yellow indicates CD-63, magenta indicates CD-81. Imaris puncta analysis was performed on BTX colocalization channels with CD-63 and CD-81. Scale bar: 10 µm. Experiment performed once. e ) Representative max intensity projected (MIP) low magnification images (upper panel) and high magnification images of BTX-positive region (lower panel) of CD63-pHluorin signal in cultured primary muscle before (pre) and immediately after application of ammonium chloride (NH 4 Cl). Dynamic magenta-orange-white indicates CD63-pHluorin intensity in low magnification images. Lower panel: magenta indicates BTX, green indicates CD63-pHluorin signal. Scale bars: 50 µm; 10 µm in upper and lower panels respectively. Experiment repeated in 2 neuromuscular co-cultures.

Article Snippet: ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle.

Techniques: Cell Culture

Journal: Nature Neuroscience

Article Title: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models

doi: 10.1038/s41593-025-02062-6

Figure Lengend Snippet: a ) Representative images of a 6DIV primary motor neuron culture infected with lentiviral (pLV) miR-126 GFP. Green indicates GFP. Scale bar: 100 µm. Repeated in 3 neuronal cultures. b ) Quantitative RT-PCR for multiple predicted and non-predicted mouse mRNA targets of miR-126a-5p in primary motor neurons expressing LV-GFP versus LV-miR126-GFP. Data are shown as the mean relative expression of each target in LV-miR126-GFP over LV-GFP ± SEM. Two-way ANOVA with Holm-Sidak correction for multiple comparisons. n = 3 neuronal cultures per condition. p = 0.0003(TDP-43), p = 0.0286(C9orf72), p = 8.78 × 10 −5 (Caspase-3), p = 0.012(NRP1), p = 0.0003(DLK1), p = 0.0016(JNK3), p = 0.7869(FUS1), p = 0.0017(CRMP4), p = 0.999(SOD1), p = 0.999(G3BP1), p = 0.9346(Sema3A), p = 0.999(COX4), p = 0.992(COX7). c ) Graphical illustrations of the human TDP-43 mRNA and two more predicted isoforms. Green indicates 3’UTRs, red marks the putative miR-126-5p target sites. d ) Representative low magnification image of healthy IPSC-derived motor neurons (IPSC-MN; KOLF) infected with pLV-hSyn-miR126. Red indicates GFP in infected neurons. Scale bar: 150 µm. image e ) Quantitative Taqman RT-PCR for miR-126-5p in human IPSC-MN (KOLF) expressing either pLV-Syn-miR126-GFP or pLV-Syn-GFP. Data are shown as the mean relative miR-126-5p expression ± SEM in miR-126 infected IPSC-derived motor neurons versus GFP infected ones. Two-tailed unpaired student’s t-test. n = 3 seperatly grown cultures. *p = 0.0162. f ) Quantitative RT-PCR for human TDP-43 mRNA with primers targeting the coding sequence (CDS) in IPSC-derived motor neurons following 7 days of pLV-Syn-miR126 or pLV-Syn-GFP infection. Data are shown as the mean relative mRNA levels of human TDP-43 mRNA ± SEM. Two-tailed unpaired student’s t-test. n = 3 separately grown cultures. **p < 0.0042. g ) Quantitative TaqMan RT-PCR for miR-126-5p in p290 SOD1 G37R GC muscles versus p290 SOD1 G37R SC. Data are shown as the mean relative level of miR-126-5p ± SEM. U6 was used as endogenous loading control. Two-tailed unpaired t-test. n = 5 mice in each group. *p = 0.0263(GC), p = 0.888(SC). h-i ) Representative images ( h ) and quantification ( i ) of miR-126a-5p FISH in NMJs of p60 B6SJL.SOD1 G93A EDL muscles versus littermate controls. Blue indicates nuclei (DAPI), green indicates NFH, magenta indicates miR-126a-5p FISH. Scale bar: 10 µm. Data are shown in violin distribution plots of the miR-126a-5p puncta per NMJ with markings of first, median and third quartiles. Two-tailed unpaired student’s t-test. n = 15; 33 NMJs from 3 mice in each condition. ****p = 3.689 × 10 −7 .

Article Snippet: ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle.

Techniques: Infection, Quantitative RT-PCR, Expressing, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Two Tailed Test, Sequencing, Muscles, Control

Journal: Nature Neuroscience

Article Title: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models

doi: 10.1038/s41593-025-02062-6

Figure Lengend Snippet: a , RT-qPCR for TDP-43 mRNA in primary muscles 5 days after miR126i treatment. GAPDH was used as loading control. RQ, relative quantification. n = 3 cultures * P = 0.0203. b , Illustration of experimental setup in c – k . Neuromuscular compartment in co-cultures was exclusively transfected at day 5 with miR126i. c , d , Representative images and analysis of TDP-43 density (puncta/100 µm) in axons within the neuromuscular compartment in control versus miR126i-treated co-cultures 5 days after transfection. Scale bar, 5 µm. n = 222, 216 axons. **** P < 1 × 10 −15 . e , f , Representative images and quantification of TDP-43 puro-PLA signal in presynaptic axons in control and miR126i-treated co-cultures. White indicates TDP-43 puro-PLA–NFH co-localization (right). Scale bar, 10 µm. n = 62, 80 NMJs. **** P = 3.39 × 10 −13 . g , h , Representative images and quantification of OPP labeling in control and miR126i-treated co-cultures. White indicates OPP–NFH co-localization (right). Scale bar, 10 µm. n = 259, 212 NMJs **** P = 1.12 × 10 −7 . i , Percent of contracting innervated muscles in control and miR126i-treated co-cultures. n = 13, 15 co-cultures. *** P = 0.00014. j , k , Representative image and quantification of NMJ disruption in control and miR126i-treated co-cultures. Scale bar, 10 µm. n = 13, 15 microfluidic co-cultures. ** P = 0.0084. l , m , Representative images and quantification of axonal TDP-43 density in control-siRNA-treated or TDP-43-siRNA-treated co-cultures in presence/absence of miR126i. Scale bar, 10 µm. n = 96, 89, 70, 73 axons. Control siRNA versus miR126i+control siRNA: **** P = 4.15 × 10 −10 ; miR126i+control siRNA versus miR126i+TDP-43 siRNA: **** P = 1.05 × 10 −7 . n , Representative images of NMJs in control-siRNA-treated or TDP-43-siRNA-treated co-cultures in presence/absence of miR126i. Scale bar, 10 µm. o , Percent of axon degeneration in co-cultures treated as described above. n = 3 co-cultures. Control siRNA versus miR126i+control siRNA: * P = 0.0296; miR126i+control siRNA versus miR126i+TDP-43 siRNA: * P = 0.0296. For h , I , data are shown as the mean ± s.d., repeated in three independent repeats. For a , k , o , data are shown as the mean ± s.e.m., repeated in three independent repeats. For d , f , m , data are shown in violin density plots with markings of first, median and third quartiles, repeated in three independent repeats. For a , d , f , h , i , k , two-tailed unpaired Student’s t -test. For m , o , one-way ANOVA with Holm–Sidak correction for multiple comparisons. In e – o , miR126i or siRNA was administered exclusively to the neuromuscular compartment. coloc, co-localization.

Article Snippet: ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle.

Techniques: Quantitative RT-PCR, Muscles, Control, Quantitative Proteomics, Transfection, Labeling, Disruption, Two Tailed Test

Journal: Nature Neuroscience

Article Title: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models

doi: 10.1038/s41593-025-02062-6

Figure Lengend Snippet: a ) Representative images from the distal (NMJ) compartment IPSCC-derived co-culture of healthy (KOLF) motor neurons and skeletal muscles. Blue indicates NFH, gray indicates BTX, magenta indicates Titin. White dashed line indicates NMJ region shown in inset. Scale bar: 20 µm. b-c ) Quantitative analysis of axonal pTDP-43 fraction ( b ) and axonal degeneration index ( c ) in iPSC-derived co-cultures of SOD1 A5V , TDP-43 M337V , and their isogenic control (KOLF). Data in ( b ) are shown in violin density plots of pTDP-43 puncta area out of NFH area of axons at the NMJ compartment with marking of first, median, and third quartiles. n = 21; 26, 18 imaging fields from 3 independently grown neuromuscular co-cultures. One-way ANOVA with Holm-Sidak correction. **p = 0.0064, ****p = 4.4 × 10 −10 . Data in ( c ) are shown as the mean NFH degeneration index of axons in the NMJ compartment ± SD. n = 28; 23; 18 imaging fields from 3 neuromuscular co-cultures. One-way ANOVA with Holm-Sidak correction. ****p = 4.66 × 10 −5 (SOD1 A5V ), 1.96 × 10 −5 (TDP43 M337V ). d ) Representative image for lentivirus-infected IPSC-MN and IPSC-muscles in co-culture within MFC. Green in IPSC-MN (upper compartment) indicates pLV-hSyn-miR126-GFP, Green in IPSC-muscles (lower compartment) indicates pLV-CMV-miR126-GFP. Scale bar: 150 µm. e ) Representative immunofluorescent images for pTDP-43 nuclear localization in SOD1 A5V and TDP-43 M337V IPSC-MN in the proximal MFC compartment of co-cultures infected with either pLV-hSyn-GFP or pLV-hSyn-miR126-GFP. Blue indicates NFH, gray indicates pTDP-43. White dashed square marks the IPSC-MN magnified in the inset. Scale bar: 20 µm. For a,d,e experiments repeated in 3 neuromuscular co-cultures.

Article Snippet: ****p = 7.68 × 10 −5 . n = 20 muscle fibers from 3 mice d ) Representative images and 3D Imaris puncta colocalization analysis of extracellular vesicle machinery in postsynaptic apparatus of enzymatically denervated EDL muscle.

Techniques: Derivative Assay, Co-Culture Assay, Muscles, Control, Imaging, Infection