Figure 8: CRISPR/CAS9-mediated disruption of mouse Fbxo22.

(a) Representative Fbxo22 wild-type (+/+), heterozygous (+/−) or nullizygous (−/−) mice at 24 weeks of age. (b) Lysates of tissues from the spleen, thymus, kidney, liver, lung and brain of 24-week-old Fbxo22 wild-type (+/+), heterozygous (+/−) or nullizygous (−/−) mice were analysed by immunoblotting using the indicated antibodies. (c) Genotype analysis of Fbxo22 wild-type (+/+), heterozygous (+/−) and nullizygous (−/−) mice by PCR. (d) Primary MEFs from Fbxo22^−/− mice showed severe growth retardation. Relative numbers of primary MEFs from Fbxo22^+/+, Fbxo22^+/− and Fbxo22^−/− mice were determined at the indicated times. Data are presented as means±s.d. of at least three independent experiments. (e) Cell cycle distribution of primary MEFs as in d was determined at 4 days by FACScan. (f) Primary MEFs from Fbxo22^−/− mice showed marked accumulations of p53, p21 and Mdm2. Primary MEFs from mice with the indicated genotypes were collected at the indicated times and the lysates were subjected to immunoblotting as shown. (g) Primary MEFs with the indicated genotypes were treated with MG132 (10 μg ml⁻¹) for 2 h. The lysates were immunoprecipitated using a control IgG, methyl (K370me2)-specific anti-p53 or anti-p53 antibodies. The resultant immunoprecipitates and lysates (Input) were subjected to immunoblotting using the indicated antibodies.