Immuno-histochemical characterization of the astrocytes derived from12 V-Ha-Ras transgenic mice. A. Upper panel shows schematic representation of the vector for the transgenic mice. Lower panel shows the presence of transgene in 12 V-Ha-Ras transgenic mice (left panel) and primary astrocytes (passage 8) derived from the V12 Ras transgenic mice (right panel) as confirmed by genotyping. Lane 2 and 4 n right panel shows negative and positive control respectively. B. Histochemical staining for LacZ in the astrocytes derived from 12 V-Ha-Ras transgenic mice. Derivative astrocytes from 12 V-Ha-Ras transgenic mice (passage 8) were stained for LacZ as a transgene marker using X-gal. Cells show nuclear positivity for LacZ (photomicrograph b). LacZ stained nuclei show prominent nuclear atypia (black arrowheads in the photomicrograph b) and dividing nucleus (white arrowhead in the photomicrograph b). U87MG glioma cells served as negative control (photomicrograph a). C & D. Immunostaining for GFAP (C) and PCNA (D) in the derivative astrocytes from 12 V-Ha-Ras transgenic mice as described in Materials and methods. Derivative astrocytes cells show cytoplasmic positivity for GFAP or nuclear positivity for PCNA (photomicrograph c) in C & D. U87MG glioma cells are used as negative control (stained without primary antibody) and positive control (stained with primary antibody) as shown in photomicrograph b and photomicrograph d respectively.