Fig. 4. Suppression of BMP2-induced osteogenesis by myogenin. MMCs were transfected with the myogenin expression plasmid pISEmgn or the control vector pISE. (A) Expression of myogenin was induced by ponasterone A simultaneously with the onset of stimulation with BMP2 for 30 hours. The expression of Runx2 was determined by immunofluorescence analysis in 132 or 177 GFP-expressing cells transfected with pISE or pISEmgn, respectively (A). MMCs transfected with pISE (B) or pISEmgn (C) were probed with antibodies to GFP and to Runx2, and immune complexes were detected by FITC or Cy3 fluorescence, respectively. Runx2 (red) was present in nuclei, whereas GFP (green) was localized to both nuclei and cytoplasm. Intrinsic fluorescence of GFP was severely decreased after the immunostaining procedure. Scale bar: 20 µm.
