fst primary antibody  (Santa Cruz Biotechnology)

Journal: Cell Transplantation

Article Title: Overexpression of Follistatin in Human Myoblasts Increases Their Proliferation and Differentiation, and Improves the Graft Success in SCID Mice

doi: 10.3727/096368909x470865

Figure Lengend Snippet: Figure 1. Overexpression of follistatin in 293T cells transfected with human follistatin lentivirus vector. (a) The human follistatin lentivirus vector was constructed by replacing the eGFP sequence in the control vector by the sequence of the short form of the human follistatin obtained from an expression vector. (b, top) 293T cells expressing the eGFP after the transfection and before the fixation (50×). (b, middle and bottom) Immunocytochemical detection of human follistatin in 293T cells transfected with pCMV- eGFP or pCMV-hFst and corresponding nuclei stained with DAPI (50×). (c) Western blot detection of follistatin in extracts of 293T cells transfected with either the eFGP or the follistatin vector.

Article Snippet: Cells were first fixed for 15 min with ethanol 95% and nonspecific binding sites Fusion Index were blocked for 1 h with PBS containing 10% FBS. pCMV-eGFP- and pCMV-hFst transduced cells were Cells were first incubated with monoclonal mouse anti- plated in a 24-well plate at 40,000 cells/well in proliferahuman follistatin primary antibody (R&D Systems, tion medium.

Techniques: Over Expression, Transfection, Plasmid Preparation, Construct, Sequencing, Control, Expressing, Staining, Western Blot

Journal: Cell Transplantation

Article Title: Overexpression of Follistatin in Human Myoblasts Increases Their Proliferation and Differentiation, and Improves the Graft Success in SCID Mice

doi: 10.3727/096368909x470865

Figure Lengend Snippet: Figure 2. Overexpression and secretion of follistatin in human myoblasts transduced with human follistatin lentivirus. (a, top) Normal human myoblasts (not fixed) expressing the eGFP after the lentivirus transduction (100×). (a, middle and bottom) Immunocytochemical detection of human follistatin in human myoblasts transduced with eGFP or with hFst lentivirus by and corresponding nuclei stained with DAPI (100×). (b) Western blot detection of human follistatin in proteins ex- tracted from transduced human myoblasts. (c) Dot blot detection of human follistatin in condi- tioned media of transduced human myoblasts.

Article Snippet: Cells were first fixed for 15 min with ethanol 95% and nonspecific binding sites Fusion Index were blocked for 1 h with PBS containing 10% FBS. pCMV-eGFP- and pCMV-hFst transduced cells were Cells were first incubated with monoclonal mouse anti- plated in a 24-well plate at 40,000 cells/well in proliferahuman follistatin primary antibody (R&D Systems, tion medium.

Techniques: Over Expression, Transduction, Expressing, Staining, Western Blot, Dot Blot

Journal: Cell Transplantation

Article Title: Overexpression of Follistatin in Human Myoblasts Increases Their Proliferation and Differentiation, and Improves the Graft Success in SCID Mice

doi: 10.3727/096368909x470865

Figure Lengend Snippet: Figure 4. Improved differentiation of normal human myoblasts transduced with a follistatin lentivirus treated or not with recombi- nant myostatin. (a) Immunocytochemical detection of myosin heavy chain (in red) on human myoblasts, incubated for 2 days in differentiation medium, expressing eGFP or hFst and treated or not with recombinant myostatin (500 ng/ml), and corresponding nuclei stained with DAPI (100×). (b) Percentage of nuclei present in MyHC-positive cells over the total number of nuclei in normal human myoblasts transduced with eGFP or hFst lentivirus, and exposed or not to 150 ng/ml of recombinant myostatin (n = 3). ϕp < 0.01; *,τp < 0.05; φp < 0.1.

Article Snippet: Cells were first fixed for 15 min with ethanol 95% and nonspecific binding sites Fusion Index were blocked for 1 h with PBS containing 10% FBS. pCMV-eGFP- and pCMV-hFst transduced cells were Cells were first incubated with monoclonal mouse anti- plated in a 24-well plate at 40,000 cells/well in proliferahuman follistatin primary antibody (R&D Systems, tion medium.

Techniques: Transduction, Incubation, Expressing, Recombinant, Staining

Journal: Cell Transplantation

Article Title: Overexpression of Follistatin in Human Myoblasts Increases Their Proliferation and Differentiation, and Improves the Graft Success in SCID Mice

doi: 10.3727/096368909x470865

Figure Lengend Snippet: Figure 5. Improved transplantation success in SCID mice of follistatin-expressing myoblasts. (a) Representative sections of SCID muscles transplanted either with pCMV-eGFP transduced my- oblasts, or pCMV-hFst transduced myoblasts and immunostained in red with an anti-human dys- trophin antibody (100×). (b) Number of human dystrophin-positive fibers in SCID mouse muscles transplanted either with pCMV-eGFP or pCMV-hFst transduced myoblasts (n = 6). *p < 0.05.

Article Snippet: Cells were first fixed for 15 min with ethanol 95% and nonspecific binding sites Fusion Index were blocked for 1 h with PBS containing 10% FBS. pCMV-eGFP- and pCMV-hFst transduced cells were Cells were first incubated with monoclonal mouse anti- plated in a 24-well plate at 40,000 cells/well in proliferahuman follistatin primary antibody (R&D Systems, tion medium.

Techniques: Transplantation Assay, Expressing, Muscles