FIG. 2.

GFP reporter expression in BAC-Kit-GFP transgenic mice. (A) RNase protection assay. Total RNA from cerebellum, testis, ovary, and liver (as indicated) of BAC200-Kit-GFP, BAC30-Kit-GFP, BAC3-Kit-GFP, and BAC200-Δ5′HS-Kit-GFP transgenic mice was processed for RNase protection assay using ³²P-labeled riboprobes specific for GFP and endogenous Kit RNA. Analysis of Kit-GFP and endogenous Kit expression in BMMC from T20, T91, and T127 BAC200-Kit-GFP mice by RNase protection and FACS analysis is shown. Mean values of fluorescence are indicated. Max, maximum. (B) Immunohistochemical detection of Kit-GFP transgene expression with anti-GFP antibody (signal in brown; hematoxylin counterstain in blue). Oocytes in the control ovary have no signal, while in the BAC200-Kit-GFP transgenic ovary, oocytes are stained in brown (red arrows). Representative sections of testis and cerebellum from nontransgenic and BAC200-Kit-GFP, BAC30-Kit-GFP, BAC3-Kit-GFP, and BAC200-Δ5′HS-Kit-GFP transgenic mice are shown. In the testis, Kit-GFP expression is found in spermatogonia (red arrow), and white stars show GFP expression in Leydig cells. The molecular layer (m) of the cerebellum shows staining in basket and stellate cell bodies as well as axons and dendrites. In addition, the molecular layer neurons form brown baskets around Purkinje cell bodies (p). Confocal microscopy identifies primordial germ cells in the mesentery and the gonadal ridge of E11.5 BAC200-Kit-GFP mouse embryos (red arrow).