Fig. 6: Soluble SIRPα does not bind to apoptotic blebs despite the presence of CD47.
From: Nanoscale imaging of CD47 informs how plasma membrane modifications shape apoptotic cell recognition
Apoptotic HeLa cells were incubated with biotinylated SIRPα and/or anti-CD47 antibody (B6H12). a SIRPα binding revealed by A647-streptavidin. b Cells were stained with anti-CD47 (B6H12) revealed by secondary antibodies labeled with Cyanine 3 (red). c Double labeling by SIRPα and anti-CD47 antibody. Cells were incubated with biotinylated human SIRPα Fc chimera protein, revealed by streptavidin A647 (red) and then fixed and stained with anti-CD47 (B6H12) revealed by secondary antibodies labeled with Alexa 488 (green). In a, b and c samples were mounted with DAPI to detect the DNA (blue) and imaged by spinning disk confocal microscopy. Z stacks of cells were acquired and the maximum projection is displayed. d Two-color STORM image showing simultaneous detection of SIRPα binding and CD47 obtained using spectral demixing (see Materials and Methods). SIRPα binding was revealed by A647-streptavidin and anti-CD47 antibody was revealed by a secondary antibody labeled with CF680. Scale bars 10 µm.
