Figure 1: Deficiency of Vilse resulted in embryonic lethality.

(a) Schematic representation of mouse vilse gene interrupted by LacZ cassette within exon 4 and 5 and two loxP sites were flanking exon 5 of Vilse gene. (b) Macroscopic inspection of embryos isolated at E13.5. ^+/+, Wild type mice; ^+/−, Heterozygous mice; ^−/−, homozygous mice. All Western blots were derived from the same cell lysates, processed in identical conditions and cropped from Supplementary Fig. S3a. GAPDH was used as a loading control. (c) Whole embryo extracts at E10.5 were lysed in RIPA buffer for immunoblotting analysis. (d) Embryos were isolated from E9.0 for hematoxylin–eosin staining. (e) In the original targeting construct, exon 1 to exon 4 of Vilse are designated to be spliced to the lacZ-trapping element. Embryos at E11.5 were stained with X-gal to detect the expression of Vilse-LacZ fusion protein. (f) Cell extracts from E9.5 embryos, and a variety of adult tissues were immunoblotted with anti-Vilse antibody. All Western blots were processed in identical conditions and cropped from Supplementary Fig. S3b. (g) Double labeling of Vilse with MAP2, Iba-1, and GFAP in the hippocampal sections of wild-type mice. DAPI indicated the position of the nucleus. Bar, 50 μm.