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Fig. 3. Redistribution of BRII in A431 cells. Colocalization of BMP receptors with caveolin-1 isoforms. A431 cells were either cultured in DMEM with 10% FBS (N) or were serum-starved (S). A431 cells cultured with serum were transfected with BRII (N +BRII) or not. After 3 days starved cells were stimulated with BMP-2 (S+B)or not stimulated (S). Cells were than fixed with paraformaldehyde-saponin (P) or acetone-methanol; BMP-receptors were labeled using polyclonal antisera against BRIa or BRII and a secondary fluorescently labeled antibody. The same cells were labeled for caveolin-1 using an antibody specific for the isoform [] or for both and ß isoforms [ß] and a secondary fluorescently labeled antibody. Cell membrane expression was visualized by confocal microscopy and 40 images were collected at the highest magnification from 40 different cells. From these images the fraction of BMP receptors colocalizing either with caveolin-1 or with and ß were calculated using ICCS. (A) The percent colocalization of BRIa with both isoforms of caveolin-1 [] (black bars) or with both isoforms (all the caveolae) as well as ß alone [ß] (white bars). (B) The percent colocalization of BRII with caveolae with both isoforms [] (black bars) and those with both isoforms and those with mainly ß [ß] (white bars).

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