myosin heavy chain mhc isoforms  (Bio-Rad)

Journal: Physiological Reports

Article Title: Macrophages modulate skeletal muscle wasting and recovery in acute lung injury in mice

doi: 10.14814/phy2.70052

Figure Lengend Snippet: Intramuscular (IM) macrophage depletion with clodronate liposomes on was performed starting on ALI day 6 with analysis on ALI day 10 (a) ( n = 9). Contralateral TA muscle was treated with vehicle control liposomes. (b) Tibialis Anterior (TA) wet weight ( n = 9). For morphometric analyses, the tibialis anterior muscle cryosections were stained for laminin‐γ1 and cross‐sectional area (CSA) was determined. The data were then expressed as (c) mean fiber CSA or as (d) distribution of CSA of the total number of myofibers analyzed. (e) The tibialis anterior muscle cryosections were also stained for MHC isoforms and expressed as mean fiber CSA for each subtype: Types IIX, IIA, IIB, IIX/A, and IIX/B. (f) Representative tibialis anterior muscle MHC stain with types IIX (black), IIA (green), IIB (red), IIX/A (intermediate green), and IIX/B (intermediate red). Values are expressed as mean ± SD. Wilcoxon signed‐rank test (paired nonparametric t test) was performed (b, c). For comparison of the distribution of fibers between two groups, a Poisson regression model was fitted in R (version 4.1.1 stats package) to predict the count of fibers with a size greater than 600 μm 2 , including the individual animal as a covariate and total number of fibers counted as an offset (d). Mann Whitney test (unpaired nonparametric t test) was performed between treatments for each fiber type (e). Figure 4a created with BioRender.com .

Article Snippet: To process samples for immunofluorescence of myosin heavy chain isoforms (Bonilla et al., ), sections were rinsed in PBST and blocked with 10% goat serum in PBS for 1 h. The mouse primary antibodies used for MHC isoforms, developed by Dr. Stefano Schiaffino (Schiaffino et al., ) and purchased through the University of Iowa Developmental Studies Hybridoma Bank, include BA‐F8 (MHC‐I, 1:50), SC‐71 (MHC‐IIa, 1:500), and BF‐F3, (MHC‐IIb, 1:100).

Techniques: Liposomes, Control, Staining, Comparison, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Small-Molecule Inhibition of MuRF1 Prevents Early Disuse-Induced Diaphragmatic Dysfunction and Atrophy

doi: 10.3390/ijms24043637

Figure Lengend Snippet: MyoMed-205 prevents early disuse-induced diaphragmatic fiber atrophy after 12 h of denervation. ( A ) Representative photomicrographs of the costal diaphragm muscle stained with hematoxylin and eosin (HE) and using immunofluorescence anti-MHC isoforms I (violet), IIa (light blue), IIb/x (black), and dystrophin (white). Scale bar = 50 μm. ( B ) Mean of diaphragm fiber types (I, IIa, and IIb/x) cross-sectional area (CSA, μm 2 ). ( C ) Fiber type distribution in the diaphragm. Data are shown as mean ± standard error of the mean (* p < 0.05 vs. Sham12h, # p < 0.05 vs. DNV12h + VEH, n = 8).

Article Snippet: Next, diaphragm fiber typing was determined by immunofluorescence assay using the following primary antibodies anti-myosin heavy chain (MHC) isoforms (Developmental Studies Hybridoma Bank, University of Iowa): anti-MHC I (#BA-D5, mouse IgG2b, 1:800), Anti-MHC IIa (#SC-71, mouse IgG1, 1:800); and diaphragm muscle membrane was stained using anti-dystrophin antibody (SC#15376, rabbit IgG, 1:500).

Techniques: Staining, Immunofluorescence