Extended Data Fig. 4: Comparison of TDP-43 and pTDP-43 accumulation in NMJs of fast versus slow muscle fibers in SOD1 mouse models.
From: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models
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.SOD1G93A 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 SOD1G37R 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 SOD1G37R 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.SOD1G93A. 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.SOD1G93A 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 SOD1G37R 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 SOD1G37R 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 SOD1G37R 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).
