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Fig. 1. Sca-1 is expressed on a subpopulation of C2C12 cells and is removed by PIPLC. (A) Flow cytometry of proliferating C2C12 cells (Pro; green) and cells grown in differentiation media for two days (Day 2; red) demonstrated that the percentage of cells with surface expression of Sca-1 increases during differentiation, but that Sca-1-positive and -negative populations persist. Gates (vertical gray bar) were established by nonspecific PE-conjugated antibody binding in each experiment (black and gray lines). A representative profile is shown with mean values for percent Sca-1-positive and -negative cells given (n=3; *significant difference, P<0.05). (B) Immunoblot analysis of Sca-1 with specific monoclonal antibody E13-161.7 in proliferating C2C12 cells (Pro) and cells grown in differentiation media for 2 and 5 days demonstrated removal of Sca-1 protein from whole cell lysates with PIPLC (5 U/ml). ß-actin was used as a control. Representative data are shown (n=4). (C) Immunostaining of Sca-1 with monoclonal antibody E13-161.7 demonstrated stripping of Sca-1 (green) from the cell surface with PIPLC (5 U/ml). Bar, 50 µm.
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