BioMed Research International / 2010 / Article / Fig 3

Research Article

L6E9 Myoblasts Are Deficient of Myostatin and Additional TGF- Members Are Candidates to Developmentally Control Their Fiber Formation

Figure 3

(a) Semiquantitative RT-PCR analysis was carried out to detect the levels of ActRIIb and follistatin levels in untransfected L6E9 (control) and in cells stably transfected with the dnActRIIb form, treated with TSA, or alternatively transfected with a short human follistatin form. Gapdh amplification was performeds as loading control. (b) Phase contrast pictures show the morphology of L6E9 cells after delivery of a dnActRIIb form, exposure to histone deacetylases inhibitor TSA, or stable follistatin overexpression, as compared to control cells. After Giemsa staining, pictures were taken under the same magnification over a time-course of 3 days. Bar = 1 0 0 𝜇 m. (c) The graphs report the quantification of the myotube size in L6E9 cells transfected with dnActRIIb, treated with TSA or overexpressing follistatin as compared to control over a 3-day time-course differentiation. 𝑃 < . 0 5 versus control. (d) Immunoblotting was performed to detect the protein levels of myogenin, Cav-3, and MyHC in L6E9 expressing the dnActRIIb form, exposed to TSA or alternatively transfected with follistatin compared to control cells. Tubulin was used as loading control.

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