Journal: Cell Death & Disease

Article Title: The histone code reader Spin1 controls skeletal muscle development

doi: 10.1038/cddis.2017.468

Figure Lengend Snippet: Surviving Spin1 M5 mice exhibit major defects in soleus, tibialis anterior, and diaphragm. ( a,b ) Appearance ( a ) and average body weight ( b ) of Spin1 M5 and control mice ( n =5 females in each category) at 30 weeks of age. Error bars represent +S.D., ** P <0.01. ( c ) Degeneration of the soleus (SOL) muscle in adult Spin1 M5 compared with control mice exemplified at 16 weeks of age. ( d ) Hind limb muscles (gastrocnemius (GC), plantaris (PL), soleus, tibialis anterior (TA), extensor digitorum longus (EDL), and quadriceps (QC)) of Spin1 M5 and control mice at 30 weeks of age. Arrows point at the soleus embedded in gastrocnemius and plantaris, which is visible in control but degenerated in Spin1 M5 mice. ( e ) Hematoxylin & eosin (H&E) staining of gastrocnemius, soleus, TA, and EDL muscle of Spin1 M5 and control mice at 30 weeks of age. ( f ) Fiber types in glycolytic (white) or oxidative (red) parts of the TA of Spin1 M5 and control mice at 15 weeks of age (top and middle rows) observed by immunofluorescence (IF) staining. Tissue sections were stained with selective antibody directed against MHC-I (purple), MHC-IIb (cyan), MHC-IIa (red), and MHC-IIx (green). For comparison, NADH staining was included (bottom row). Fibers with abnormal NADH staining are marked with arrows. Corresponding fibers in each column of images are squared

Article Snippet: For immunofluorescence staining the following primary antibodies were used: SPIN1(5865) 1 μ g/ml; Pax7 (PAX7, DSHB, batch 7/2/15) 2 μ g/ml; Tcf4 (6H5-3, Millipore, Darmstadt, Germany, 05-511, batch 2155406) 10 μ g/ml; Ankrd1 (Proteintech Group, Manchester, UK, 11427-1-AP, batch 1951) 1:100; Ankrd2 (Proteintech Group, 11821-1-AP, batch 7649) 1:100; dystrophin (Abcam, Cambridge, UK, ab15277, batch GR226781-6) 1:500; MHC-I (NOQ7.5.4D, Sigma, Munich, Germany, M8421, batch 035M4792V) 1:2000; MHC-IIa (SC-71, DSHB, batch 4/7/16) 1:10; MHC-IIx (6H1, DSHB, batch 3/3/16) 1:6; MHC-IIb (BF-F3, DSHB, batch 5/12/16) 1:20; MCH, skeletal, fast (MY-32, Sigma, M4276, batch 083M4790V) 1:1000; GNZ (anti-GFP, Abcam, ab13970, batch GR236651) 1:1000; normal rabbit IgG (Santa Cruz, Heidelberg, Germany, sc-2027).

Techniques: Control, Muscles, Staining, Immunofluorescence, Comparison

Journal: Science Advances

Article Title: Lmod2 is necessary for effective skeletal muscle contraction

doi: 10.1126/sciadv.adk1890

Figure Lengend Snippet: Lmod2 WT + cGFP and KO + cGFP-Lmod2 skeletal muscle cross sections were stained with antibodies against different MHC isoforms. Representative images of EDL ( A ) or soleus ( B ) MHC fiber type staining. Top: MHC type I (blue), MHC type IIB (green), and laminin (red). Middle: MHC type IIA (green) and laminin (red). Bottom: MHC type IIX (blue) and laminin (red). Quantification of the number of MHC type I–, IIA–, IIB–, or IIX–positive fibers in EDL ( C ) or soleus ( E ) cross-sections shown as percent of total fibers. Minimum Feret diameter of MHC type I–, IIA–, IIB–, or IIX–positive fibers in EDL ( D ) or soleus ( F ) cross sections. Scale bars, 100 μm. All values are means ± SD; P < 0.05 was considered significant, unpaired t test; * P < 0.05, ** P < 0.01; n = 4 mice per genotype.

Article Snippet: The following primary antibodies were used: MHC I [3.5 mg/ml; BA-F8 IgG2b, Developmental Studies Hybridoma Bank (DSHB)], MHC IIB (9 mg/ml; BF-F3 IgM, DSHB), MHC IIA (3.5 mg/ml; SC-71 IgG1, DSHB), MHC IIX exclusion (3.5 mg/ml; BF-35 IgG1, DSHB), pan-myosin (9 mg/ml; A4-1025 IgG2a, DSHB), and rabbit polyclonal laminin (1:400; L9393, Sigma-Aldrich).

Techniques: Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Cistanche tubulosa (Schenk) Wight Extract Enhances Hindlimb Performance and Attenuates Myosin Heavy Chain IId/IIx Expression in Cast-Immobilized Mice

doi: 10.1155/2019/9283171

Figure Lengend Snippet: MyHC IId/IIx protein quantification in the TS muscle in each group (a)–(c). (a) Western blot of MyHC IId/IIx and GAPDH. (b) Densitometric measurements of MyHC IId/IIx at the end of the experiment. (c) Densitometric measurements of GAPDH (loading control) at the end of the experiment. The Smirnov–Grubb's test was used to exclude outliers and one-way ANOVA for analysis. Data are shown as mean ± SEM ( n = 5–6). ∗∗ p < 0.01 vs. cast (+)/vehicle group.

Article Snippet: The antibodies were chosen and diluted as follows: mouse anti-MyHC IId/IIx monoclonal antibody (1 : 1000, clone A4.1025, #05-716, Merck Millipore, Burlington, Massachusetts, United States), mouse anti-GAPDH loading control monoclonal antibody (1 : 3500, G041 abm, Vancouver, Canada), and HRP-conjugated goat antimouse IgG polyclonal secondary antibody H&L (1 : 2000, Abcam, Cambridge, United Kingdom).

Techniques: Western Blot