Journal: The FEBS journal

Article Title: GPR56 promotes myoblast fusion through SRE- and NFAT-mediated signaling but is not essential for muscle development in vivo

doi: 10.1111/febs.12529

Figure Lengend Snippet: A. H&E staining of GPR56 WT and KO gastrocnemius muscle at days 4, 6, and 18 after cardiotoxin injury. KO morphology and timing does not look different from WT. B. mRNA expression of GPR56 by RT-qPCR shows transient upregulation of GPR56 during regeneration. C. Myofiber diameter in cardiotoxin-injured WT and GPR56 KO gastrocnemius muscle shows no difference in diameter between WT and KO. D. mRNA expression by RT-qPCR of various genes in WT (black diamond) and GPR56 KO (gray circle) cardiotoxin-injured muscle. Myf5, MyoD, and myogenin are delayed in expression in KO muscle. * p<0.05, n = 3. E. Sample Western blots of myosin heavy chain protein expression. F. Quantification of the amount of MHC I, IIA, or IIB protein expression by Western blot in WT and KO gastrocnemius muscle in mice of various ages shows no difference in the amount of MHC isoforms between WT and KO. G. Quantification of the % of positive MHC I, IIA, or IIB fiber types in KO versus WT muscles, based on immunofluorescence staining in 4 littermate pairs.

Article Snippet: The antibodies against MHC Type IIA and MHC Type IIB, developed by S. Schiaffino, were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242.

Techniques: Staining, Expressing, Quantitative RT-PCR, Western Blot, Muscles, Immunofluorescence

Journal: Skeletal Muscle

Article Title: Elusive sources of variability of dystrophin rescue by exon skipping

doi: 10.1186/s13395-015-0070-6

Figure Lengend Snippet: Variability of dystrophin protein expression, as shown by IF after PMO injection. a Representative images of C57BL/10 (WT) and b PMO-treated mdx tibialis anterior sections stained for dystrophin. The WT control shows uniform IF staining for dystrophin. Insert at ×40 shows expected staining pattern for dystrophin-positive fibers. b PMO-treated mdx tibialis anterior shows a mosaic staining pattern and clustering of positive fibers. The yellow line represents the border between the tibialis anterior and EDL. Quantification was performed on the entire area of the muscle section. c Representative images of mouse mdx-6 showing variability between the muscles of the same animal. Images were selected to show positive fiber clustering and do not represent total area quantification. d IF quantification of diaphragm, gastrocnemius, heart, quadriceps, tibialis anterior, and triceps for all mice ( n = 6). e Geographic variability observed within the highly rescued triceps from mouse mdx-1. All tissues were sectioned (10-μm thick), stained, and probed with goat anti-rabbit IgG Alexa 594 antibody. Dystrophin-positive fibers were normalized to the area of the muscle section and the WT percentage of positive fibers. Original magnification for a, b, e = ×20; scale bar , 500 μm; for c = ×40; scale bar , 100 μm

Article Snippet: Muscle fiber types were identified using the following antibodies: mouse IgG2b monoclonal anti-type 1 MHC (clone BA-D5, 1:50), mouse IgG1 monoclonal anti-type 2a MHC (clone SC-71, 1:50), mouse IgM monoclonal anti-type 2b MHC (clone BF-F3, 1:5), and mouse IgG1 monoclonal anti-embryonic MHC (clone F1.652, 1:25), all obtained from the Developmental Studies Hybridoma Bank at the University of Iowa (Ames, IA, USA) [ ].

Techniques: Expressing, Injection, Staining, Control, Muscles

Journal: The Journal of Cell Biology

Article Title: Different modes of hypertrophy in skeletal muscle fibers

doi: 10.1083/jcb.200105147

Figure Lengend Snippet: Trend in fiber type changes in gracilis anterior and posterior muscle influenced by exercise not transgene. (A) Gracilis anterior muscle showed a trend of increasing type IIB MyHC at the cost of type IIA/X MyHC–positive fibers. A significant difference was only found in WT-EX and transgenic mice. (B) Fiber type changes in gracilis posterior muscle showed a trend toward type IIA/X MyHC–positive fibers at the cost of fibers expressing type IIB and type I MyHC. Exercise appears to be the main influence on these trends.

Article Snippet: The sections were incubated with either anti-MyHC I antibody (American Type Culture Collection A4.951; 1:10 dilution in PBS/1% BSA) or anti-MyHC IIB antibody (American Type Culture Collection BF-F3; 1:10 dilution in PBS/1% BSA), washed three times in PBS, and incubated in secondary antibody (Jackson ImmunoResearch Laboratory; peroxidase-conjugated goat anti–mouse IgG 115-035-164, or peroxidase-conjugated goat anti–mouse IgM 115-035-075, both 1:500 in PBS/1% BSA).

Techniques: Transgenic Assay, Expressing