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. 2013 Jul 9;288(34):24528–24539. doi: 10.1074/jbc.M113.484014

FIGURE 1.

FIGURE 1.

Targeted disruption of the Stra6 gene. A, structure of the targeting vector and partial restriction maps of the WT Stra6 locus before ((+) allele) and after (L3 allele) homologous recombination, as well as after FLPe- and Cre-mediated excision (L2 and L− alleles, respectively). Numbered black boxes indicate exons. The locations of restriction sites (A, AijI; B, BclI; X, XbaI) are indicated. Arrowhead flags represent loxP sites; neo, FRT-flanked neomycin resistance cassette. Arrows indicate the location and orientation of PCR primers 1–3. B, PCR analysis of tail genomic DNA from mice with the indicated genotype using primers 1 and 2 (upper panel) and 1–3 (lower panel). The identities of the different alleles are indicated on the right. C, left panel, immunoblots of STRA6 in protein extracts from E9.5 embryos with the indicated Stra6 genotype, attesting for the lack of STRA6 in the mutant sample. Control for protein loading was assessed using anti-actin antibodies (ACT). Right panel, immunoblot of STRA6 in lysates from COS-7 cells transfected with plasmids driving expression of either STRA6 or RBPR2, as indicated, attesting for the specificity of the anti-STRA6 antibody. Controls for protein loading and transfection efficiency were assessed using anti-tubulin (TUB) and anti-β-gal antibodies, respectively. D–I, immunodetection of STRA6 (red signal) on histological sections from WT (D, F, and H) and Stra6-null (E, G, and I) choroid plexuses (D and E), eyes (F and G), and testes (H and I). Cell nuclei were counterstained with DAPI (H and I). Note that the faint signals detected in the center of the seminiferous tubules at stages II–VI and VII–VIII in control (H) and in mutant (I) testes correspond to an unspecific staining of elongated spermatids. CP, choroid plexus; RPE, retinal pigment epithelium; SC, Sertoli cell; NR, neural retina. The dotted line (E) encircles a choroid plexus. Brackets (F and G) encompass the neural retinal layers. Roman numerals (H and I) indicate stages of the seminiferous epithelium cycle. Scale bar is shown in I and is 150 μm for D and E, 110 μm for F and G, and 100 μm for H and I.