Journal: Journal of Biomedicine and Biotechnology

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

doi: 10.1155/2010/326909

Figure Lengend Snippet: (a) Morphological visualization of C2C12 and L6E9 myoblasts undergoing differentiation. After Giemsa staining, phase contrast pictures were taken under the same magnification. Bar = 100 μ m. (b) The graph represents the quantification of myotube average size in differentiating C2C12 and L6E9 cells. * P < .05. (c) Immunoblotting was performed to compare the time-course expression of myogenin, Cav-3 and MyHC between C2C12 and L6E9 cells. Tubulin was used as loading control. (d) Semiquantitative RT-PCR analysis was performed to detect the transcript levels of myostatin, follistatin, ActRIIa, and ActRIIb in C2C12 and L6E9 cells cultured in differentiating medium. Gapdh amplification was performed as loading control. (e) RT-PCR analysis was carried out to amplify a 377 bp long fragment of myostatin in both mouse and rat gastrocnemius muscles. The amplification was performed by using the total RNA processed in presence or absence of reverse transcriptase (RT+ or RT−, resp.). Total RNAs are shown as loading control.

Article Snippet: The following primary mouse monoclonal antibodies (1 : 1000 diluted) were used: anti-Myogenin (clone F5D, Santa Cruz Biotechnology), anti-MyHC (Hybridoma Bank, University of Iowa), and anti-Cav-3 (clone 26, BD Transduction Laboratories).

Techniques: Staining, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Amplification

Journal: Journal of Biomedicine and Biotechnology

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

doi: 10.1155/2010/326909

Figure Lengend Snippet: (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 = 100 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 < .05 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.

Article Snippet: The following primary mouse monoclonal antibodies (1 : 1000 diluted) were used: anti-Myogenin (clone F5D, Santa Cruz Biotechnology), anti-MyHC (Hybridoma Bank, University of Iowa), and anti-Cav-3 (clone 26, BD Transduction Laboratories).

Techniques: Reverse Transcription Polymerase Chain Reaction, Stable Transfection, Transfection, Amplification, Over Expression, Staining, Western Blot, Expressing