Fig. 6: Generation of Thoc6^fs/fs mouse model.

A CRISPR/Cas9 editing strategy to introduce frameshift variants in mouse Thoc6. sgRNA sequence highlighted in red text. B Representative images of isolated Thoc6^+/+, Thoc6^fs/+, and Thoc6^fs/fs whole embryos at E9.5. Scale bar: 50 μm. C Western blot analysis with quantifications of THOC6 levels in E8.5 and E9.5 mouse embryos. β-actin, loading control. Genotypes: Thoc6^+/+ (black), Thoc6^fs/+ (dark red), Thoc6^fs/fs (pink). Data shown as mean ± SEM. Significance, two-tailed t test, **** indicates p = <0.0001. This experiment was repeated three times independently and showed similar results. D Litter ratio analysis for E8.5–9.5 (left) and weaned (right) Thoc6^fs/fs mice. Ratios are consistent with embryonic lethality of homozygous frameshift mice. Genotypes: Thoc6^+/+ (black) Thoc6^fs/+ (dark red), Thoc6^fs/fs (pink). N = 164 mice (left); n = 27 mice (right). E Immunostaining of markers PH3 and C.CASP3 in E9.5 mouse forebrain. Illustration highlights sectioning and quantification approach. 60x magnification; Scale bar: 25 μm. F Quantifications of fractions of PAX6, PH3, and C.CASP3-expressing cells in E9.5 neuroepithelium. Measurements were combined from one rostral and one caudal section (from two lateral segments depicted by solid black boxes in E). N = 3 embryo replicates per genotype. Genotypes: Thoc6^+/+ (black) and Thoc6^fs/fs (pink). Data shown as mean ± SEM. Significance, two-tailed t test. Source data are provided as a Source Data file.