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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.

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