Fig. 5. Deletion of Ring1 activates BMP and Wnt signaling in the early stage telencephalon.

a–c, e, f, h–j Coronal sections of the brain of Ring1A KO or Ring1A/B dKO mice at E10 were subjected to in situ hybridization analysis of Wnt8b (a–c), Axin2 (e, f), or Bmp4 (h–j) mRNA with the use of RNAscope⁵⁴. The regions indicated with outlined squares (200 by 200 μm in a and h, 300 by 300 μm in e, 40 by 40 μm in b and i) are shown at higher magnification in (b), (f), (I), (c), and (j), respectively. The telencephalic wall is demarcated by dotted lines. Arrowheads in (a) represent the ventral border of Wnt8b^high region. Scale bars, 200 μm. d, g Quantification of intensity of Wnt8b and Axin2 mRNA for images similar to those in (a) and (e), respectively. The telencephalic wall was divided into 10 bins, from 1 (dorsal) to 10 (ventral), and the average intensity was determined in each bin and normalized by the average value for bin 1. Data are means ± s.d., n = 3 embryos of each genotype. k The average density of Bmp4 signals (per 10⁴ μm²) of the dorsal, ventral, and ventralmost regions of the Ring1A KO and Ring1A/B dKO mouse telencephalon was determined for images as in (h). Data are means ± s.d., n = 3 embryos of each genotype. l, m Coronal sections of the brain of Ring1A KO or Ring1A/B dKO mice at E10 were subjected to immunohistofluorescence staining with antibodies to Id1. The regions indicated with yellow outlined rectangles (150 by 200 μm) in (l) are shown at higher magnification in (m). Nuclei were counterstained with Hoechst 33342. The telencephalic wall is outlined by yellow dotted lines in (m). Scale bars, 200 μm. n The average intensity of Id1 signals in the ventral 90% of the telencephalic wall normalized by that for the dorsal 10% (dorsal midline) was determined from images similar to those in (l). Data are means ± s.d., n = 3 embryos of each genotype. Two-tailed Student’s unpaired t test. Source data are provided as a Source Data file.