Fig. 3. Effect of the Gcm genes in cultured brain cells. (A-G) Primary cultured cells from E12 mouse hemispheres were transduced with control (A,D), mouse Gcm1 (B,E,G) or mouse Gcm2 (C,F) retrovirus and cultured in chemically defined medium for 3 days (A-F) or 2 days (G). The cells were stained with X-Gal, followed by staining with anti-GFAP (A-C,G) or anti-MAP2 (D-F) antibodies. GFAP⁺ (H) or MAP2⁺ (I) cells in X-gal⁺ cells after 3 days culture were counted. (J-N) After control (J,K) or mouse Gcm1 (L,M) transduction, brain cells were stained with both ß-galactosidase (red) and S100ß (green) antibodies. J and L are merged images. Arrowheads show overlapped signals in mouse Gcm1 transduction. The percentage of overlap was plotted in N. (O,P) Brain cells were infected with control lacZ or mouse Gcm1-lacZ virus together with virus harboring alkaline phosphatase (ALK). lacZ⁺ and ALK⁺ cells were visualized by X-gal-staining (arrows) and NBT-staining (arrowheads), respectively. They were further stained with anti-GFAP antibodies (brown). Cells double-stained for X-gal and anti-GFAP Ab or ALK and anti-GFAP Ab were counted (P). Scale bars: 30 µm in A-G; 40 µm in J-M,O.