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. 2004 May 10;75(1):106–111. doi: 10.1086/422103

Figure 3.

Figure 3

Western immunoblotting was performed under reducing and nonreducing conditions (with or without β-mercaptoetanol [β-ME]) by use of anti-His-HRP antibody (Roche) or anti-Myc antibody (Invitrogen), as appropriate. Immunofluorescent signals were detected with the use of the ECL system (Amersham). Western blots performed with anti-Myc antibody are shown. A, Analysis performed in the presence of β-ME. Bands corresponding to mature (duplet at ∼21–23 kDa) and precursor (at ∼50–55 kDa) BMP15-Myc-His chimeras were seen in the WT and mutant (MUT) lanes but were not seen in the control media from nontransfected (NT) cells. B, Analysis performed under nonreducing conditions allowing the visualization of dimeric products. Bands corresponding to mature (∼45 kDa) and precursor (>110 kDa) dimers were seen in both WT and MUT lanes. Additional bands corresponding to precursor monomers (∼50–55 kDa) or to stable precursor-mature dimers (∼80 kDa) were detected in the mutant Y235C-BMP15 but not in WT lanes, even when double the amount of protein was tested (WT 2x). The band at 65–70 kDa in both panels corresponds to a nonspecific cross-reacting protein present in all media.