Fig. 3. SFRP1 affects cell differentiation in retina cell cultures. E5 chick retinas were dissociated and grown for 24 hours in the presence of CM from (A,C,E) MDCK/control or (B,D,F) MDCK/SFRP1. After fixation, cultures immuno-stained with antibodies against (A,B) anti-NF, (C,D) islet1/2, (E,F) visinin or phospho-histone3 (PH3, a mitotic marker) and counterstained with Hoescht (blue staining). (G) Final cell density (Hoescht⁺), the degree of differentiation (Tuj1⁺; islet1/2⁺; visinin⁺) and mitotic rate (PH3⁺) were evaluated in control (empty bars) and treated cultures (filled bars) by counting, in each experimental condition, the number of immuno-positive cells and the total number of cells in 14 randomly selected microscopic fields. Data represent a typical experiment performed in triplicate. Note how the presence of SFRP1 increases the number of differentiated cells without significant variations in the total number of cells or in the rate of cell division. *P<0.05; ***P<0.0001, Student's t-test. Bar, 185 µm.

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