Journal: Cell reports

Article Title: Misregulation of Drosophila Myc Disrupts Circadian Behavior and Metabolism

doi: 10.1016/j.celrep.2019.10.022

Figure Lengend Snippet:

Article Snippet: Primary antibodies used include: mouse anti-dMyc (from Eisenman lab), rabbit anti-α-Tubulin (Cell Signaling, Danvers, MA, USA) and Guinea Pig anti-PER (GP1140).

Techniques: Recombinant, Western Blot, Reverse Transcription, Lysis, Marker, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Software

Journal: EMBO Molecular Medicine

Article Title: Apoptosis inhibitors and mini-agrin have additive benefits in congenital muscular dystrophy mice

doi: 10.1002/emmm.201100151

Figure Lengend Snippet: A. Percentage of CNF. In dy W / dy W mice, only 9% of the muscle fibres are centrally nucleated, indicative of impaired regeneration. In dy W /mag muscle the number of CNF is increased and is further elevated by co-expression of Bcl2 in both 12- and 16-week-old dy W /Bcl/mag mice (two-way ANOVA over all ages: p < 0.0001); N = 4. B. Spontaneous muscle regeneration in 12-week-old muscles. Antibodies to dMyHC (green) and laminin-γ1 (red) were used. DAPI (blue) visualizes nuclei. Only few dMyHC-positive fibres are found in dy W / dy W mice. Some more dMyHC-positive fibres are present in dy W /mag (arrows) muscle, whereas, many more are detected upon Bcl2 expression in dy W /Bcl and dy W /Bcl/mag (encircled areas). C. Quantification of dMyHC-positive fibres. The number of dMyHC-positive fibres is highest in Bcl2 transgenic mice ( dy W /Bcl and dy W /Bcl/mag) followed by dy W /mag and dy W / dy W mice. Although not significant at 12 or 16 weeks of age ( t -test), the increase of dMyHC-positive fibres in dy W /Bcl/mag compared to dy W /mag muscles becomes significant when measured over all ages (two-way ANOVA: p = 0.022). No spontaneous regeneration was detected in control muscles (ctrl); N ≥ 4 ≤ 6. D.–F. Regenerative response of tibialis anterior muscle of 6-week-old mice after notexin injection. D. Muscle cross-sections were stained with antibodies to dMyHC (green) and laminin-γ1 (red), and with the nuclear marker DAPI (blue) 1 week after notexin injection. Only very few dMyHC-positive fibres are detected in dy W / dy W mice, while many muscle fibres are dMyHC-positive in dy W /Bcl, dy W /mag, dy W /Bcl/mag and control (ctrl) mice. Note that muscle fibres in dy W /Bcl mice are small; N ≥ 4 ≤ 5. E. Regeneration status of tibialis anterior 2 weeks after notexin injection. Antibodies to dMyHC (green), laminin-γ1 (red) and the nuclear marker DAPI (blue) were used. Absence of dMyHC-positive fibres and large areas devoid of laminin-γ1 staining (asterisks) are indicators of the failed regeneration in d y W / dy W and dy W /Bcl muscles. The presence of few dMyHC-positive fibres and the non-disrupted laminin-γ1 staining indicate successful muscle regeneration in dy W /mag, dy W /Bcl/mag and control mice; N ≥ 3 ≤ 4. F. H & E staining of tibialis anterior 2 weeks after notexin injection shows the failure in regeneration leading to fibrosis in dy W / dy W and dy W /Bcl muscles (asterisks) and the successful regeneration in dy W /mag, dy W /Bcl/mag and control mice; N ≥ 3 ≤ 4. All values represent the mean ± SEM; N indicates the animal number per each experimental group. p -Values are Student's t -test (*** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05; n.s. p > 0.05) or two-way ANOVA as noted in the text. Size bars = 50 µm.

Article Snippet: The antibodies used for immunofluorescence were purchased from the following commercial sources: Monoclonal mouse anti-rat dMyHC, monoclonal rat anti-mouse laminin-γ1 chain (Chemicon, MAB1914) and monoclonal rat anti-mouse F4/80 (Abcam, ab6640).

Techniques: Expressing, Transgenic Assay, Injection, Staining, Marker

Journal: Skeletal Muscle

Article Title: Angiotensin II type 1 receptor antagonists alleviate muscle pathology in the mouse model for laminin-α2-deficient congenital muscular dystrophy (MDC1A)

doi: 10.1186/2044-5040-2-18

Figure Lengend Snippet: L-158809 improved muscle regeneration, body weight, locomotion, and muscle strength in dy W /dy W mice. (A) L-158809 ( dy W -L158) did not significantly affect the number of centrally nucleated fibers (CNF) in either triceps or diaphragm muscle of dy W /dy W mice (n ≥ 4). (B) Staining of diaphragm muscle for the regeneration marker developmental myosin heavy chain (dMyHC; green), the basement membrane marker laminin-γ1 (red), and the nuclear marker 4",6"-diamidino-2-phenylindole hydrochloride (DAPI) (blue). L-158809 enabled damaged dy W /dy W muscle fibers to regenerate, whereas in non-treated dy W /dy W muscle many dMyHC-expressing cells continued degenerating. (C) L-158809 application strongly increased the number of intact regenerating muscle fibers in dy W /dy W mice (n ≥ 4). (D) Tibialis anterior (TA) muscle stained for dMyHC (green), laminin-γ1 (red), and DAPI four days after notexin-induced injury. L-158809 increased the regenerative capacity of damaged dy W /dy W muscle. (E) The number of dMyHC-expressing fibers in TA muscle of dy W / dy W mice 5 days after notexin injection was more than 8 times higher in L-158809 treated dy W /dy W mice but was still lower than in controls (n = 3). (F) L-158809 increased the average body weight of dy W /dy W mice from 7.6 g to 9.5 g, although age-matched wild-type (WT) mice weigh more than 20 g. (n ≥ 12). (G) . L-158809 doubled the time a dy W /dy W mouse could hold itself on a vertical grid, from 26 seconds to 51 seconds (P = 0.0001) (n ≥ 14). (H) L-158809 increased the time dy W /dy W mice explored an unknown surrounding (P = 0.0004) within 10 minutes (n ≥ 12). All values represent the mean ± SEM (n ≥ 4). One-way ANOVA: ** P ≤ 0.001; * P ≤ 0.05; n.s. (non-significant) P > 0.05. Scale bar = 50 μm, if not indicated differently.

Article Snippet: Monoclonal rabbit anti-mouse TGF-β (56E4, CST #3709), monoclonal rabbit anti-human phospho-Smad2(Ser465/467)/Smad3(Ser423/425) antibody (CST #9510) (both Cell Signaling Technology, Beverly, MA, USA) polyclonal rabbit anti-human TSP 1 (LS-C26356; Lifespan Biosciences, Inc., Seattle, WA, USA), polyclonal rabbit anti-human periostin (RD181045050; BioVendor LLC, Candler, NC, USA), monoclonal rat anti-mouse F4/80 (ab6640; Abcam, Cambridge, MA, USA), monoclonal mouse anti-rat developmental myosin heavy chain (dMyHC) (NCL-MHCd; Novocastra, Norwell, MA, USA), monoclonal rat anti-mouse laminin-γ1 chain (MAB1914; Chemicon (now EMD Millipore, Billerica, MA, USA)).

Techniques: Staining, Marker, Membrane, Expressing, Injection

Journal: The FASEB Journal

Article Title: Satellite cells deficiency and defective regeneration in dynamin 2‐related centronuclear myopathy

doi: 10.1096/fj.202001313rrr

Figure Lengend Snippet: FIGURE 5 Formation of new myofibers during regeneration after injury. New myofibers express developmental myosin heavy chain (in red). Membranes were delimited with laminin antibody (in green), nuclei were stained with DAPI (in blue). Arrows indicate dMyHC-positive fibers. Scale bar = 50 μm. A, WT and HTZ uninjured muscles. B, Formation of new myofibers after EI injury. C, Quantification of dMyHC-positive fibers in relation to the total number of fibers after EI lesion. N = 5 individuals per genotype at time point zero, n = 8 individuals per genotype after lesion, except HTZ 5 days and WT 15 days, with n = 7 each. D, Formation of new myofibers after CTX injection. E, Quantification of dMyHC- positive fibers in relation to the total number of fibers after CTX lesion. N = 5 individuals per genotype at time point zero, n = 3 individuals per genotype after lesion. In C and E, five to seven random fields were manually counted per animal

Article Snippet: Muscle cryosections were labeled with primary antibodies rabbit anti-laminin (1:300 dilution, Z0097, Dako), mouse anti-developmental myosin heavy chain (dMyHC) (1:30, NCL-MHCd, NovoCastra), anti-Ki67 (1:200, ab5580), and mouse anti-PAX7 (1:20, concentrate, Developmental Studies Hybridoma Bank), overnight at 4°C.

Techniques: Staining, Muscles, Injection

Journal: Frontiers in Neurology

Article Title: Induced degeneration and regeneration in aged muscle reduce tubular aggregates but not muscle function

doi: 10.3389/fneur.2024.1325222

Figure Lengend Snippet: Gastrocnemius muscle sections of C57BL/6J before (NE, A ) and after electroporation (5, 15, and 30 dpe, B–D ) and in Dmd mdx mice (E) . IF was performed using antibodies against dMyHC (red stain inside muscle fibers) and laminin (green stain). Nuclei are stained in blue by dapi (4’,6-diamidino-2-phenylindol). (F) Denotes the percentage of positive dMyHC fibers between experimental groups. The only significant difference was between NE C57BL/6J and 5 dpe mice (Kruskal–Wallis with Dunnett’s correction).

Article Snippet: Primary antibodies used for staining were a 1:30 dilution of mouse dMyHC-NCL, for developmental myosin staining (Novocastra NCL-MHCd, Leica Biosystems, Concord, ON, Canada) and a 1:100 dilution of rabbit anti-laminin (Z0097, Dako, San Diego, CA, United States) for extracellular matrix staining.

Techniques: Electroporation, Staining