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Fig. 7. (A-C) GFP-positive neurons express cSIRP{alpha} in co-transfected cultures. (A) GFP fluorescence of neurons; (B) the same neurons labeled with anti-cSIRP{alpha}, an anti-SIRP{alpha} raised against C-terminal peptide (Alexa Fluor® 594-conjugated secondary antibody); (C) merger of the two images. Anti-cSIRP{alpha} labels non-transfected neurons (abundant in the field) only very weakly so that they are not visible. Bar, 20 µm. (D-I) Overexpression of mutant SIRP{alpha} in the growth cone. (D-F) Representative phase-contrast images of growth cones transfected with: GFP only (D), cSIRP{alpha} plus GFP (E), or cSIRP{alpha}-FYFF plus GFP (F). (G) Distribution of endogenous SIRP{alpha} (labeled with an antibody to the external domain) in a growth cone overexpressing cSIRP{alpha}. (H) Immunolocalization of cSIRP{alpha} overexpressed in the growth cone (labeled with anti-cSIRP{alpha}). (I) The phase-contrast image of the growth cone in H. A process of a non-transfected neuron, visible only in phase-contrast, runs across the image. It is not detectable by immunofluorescence. Secondary antibodies were Alexa Fluor® 594 conjugated. Bar, 10 µm.