Frozen mouse tibialis anterior (TA) muscle from 35 day old mice was cut into 8-mm cross-sections and stained with Mayer’s haematoxylin and eosin (H&E). The number of centrally nucleated fibres and myofibers size quantification was performed manually at 40X magnification with an Olympus BX43 light microscope. Immunofluorescence (IF) was conducted as previously described [43 (link)]. For fibre typing, double IF was performed using antibodies against dystrophin (Abcam #ab15277, 1/500) in combination with either myosin heavy chain type 1 (type I, slow, Sigma #M8421, 1/50) or myosin heavy chain type 2b (fast, DSHB, #BF-F3,1/50). Myofiber size measurements were reported as minimal Feret’s diameter as previously described [53 (link)]. In addition, a Fiji macro (see Supplementary Materials Table) was written to automatically detect and quantify cross-sectional area of fibres outlined by IF staining to dystrophin (Supplementary Fig. 3 ). For p-SMAD immunofluorescence, p-SMAD2/SMAD3 (Thr8) was used (ThermoFisher, cat # PA5–99378, 1/100) and the fraction of positive p-SMAD 2/3 nuclei/total nuclei were determined using ImageJ.
P smad2 smad3 thr8
Manufactured by Thermo Fisher Scientific
The P-SMAD2/SMAD3 (Thr8) is a laboratory reagent used for the detection and quantification of phosphorylated SMAD2 and SMAD3 proteins. It is primarily used in research applications to study cellular signaling pathways involving the TGF-beta/SMAD signaling cascade.
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Lab products found in correlation
2 protocols using p smad2 smad3 thr8
1
Quantifying Muscle Fiber Characteristics
2
Histological and Molecular Analysis of Mouse Skeletal Muscle
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Frozen mouse tibialis anterior (TA) muscle from 35 day old mice was cut into 8-mm cross-sections and stained with Mayer’s haematoxylin and eosin (H&E). The number of centrally nucleated fibres and myofibers size quantification was performed manually at 40X magnification with an Olympus BX43 light microscope. Immunofluorescence (IF) was conducted as previously described [43 (link)]. For fibre typing, double IF was performed using antibodies against dystrophin (Abcam #ab15277, 1/500) in combination with either myosin heavy chain type 1 (type I, slow, Sigma #M8421, 1/50) or myosin heavy chain type 2b (fast, DSHB, #BF-F3,1/50). Myofiber size measurements were reported as minimal Feret’s diameter as previously described [53 (link)]. In addition, a Fiji macro (see Supplementary Materials Table) was written to automatically detect and quantify cross-sectional area of fibres outlined by IF staining to dystrophin (Supplementary Fig. 3). For p-SMAD immunofluorescence, p-SMAD2/SMAD3 (Thr8) was used (ThermoFisher, cat # PA5-99378, 1/100) and the fraction of positive p-SMAD 2/3 nuclei/total nuclei were determined using ImageJ.
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