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Fig. 4 | Skeletal Muscle

Fig. 4

From: High-throughput screening identifies modulators of sarcospan that stabilize muscle cells and exhibit activity in the mouse model of Duchenne muscular dystrophy

Fig. 4

OT-9 increases laminin-binding adhesion complexes at cell surface. C2C12 myotubes treated with vehicle or 5 μM of OT-9 for 48 h were incubated in amine-reactive biotin to label cell surface proteins. Avidin was used to affinity purify the labelled proteins before immunoblot analysis with a antibodies recognizing the laminin-binding glycoepitope of alpha-dystroglycan (α-DG (glycan)) or core alpha-dystroglycan. b In cells treated with OT-9, both glycosylated alpha-dystroglycan and core alpha-dystroglycan were increased at the cell surface. c–d Quantification of immunoblots. e mdx myotubes treated with vehicle or 5 μM of OT-9 for 48 h were incubated in biotin to label cell surface proteins and affinity purified with avidin. Immunoblot analysis shows upregulation of utrophin associated with biotin-labelled cell surface proteins. f Quantification of utrophin immunoblot. Data represents individual replicates and mean value. n = 3. α-DG, alpha-dystroglycan; UTRN, utrophin; R.U., relative units. *p < 0.05

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