Related to Fig. 1 : The generation and validation of Mki67 ^−/− mice deficient for Ki67. (A) Schematic of the recombining strategy to target the Mki67 locus. FRT, flippase recognition target. (B) Schematics of the Southern screening strategy used for genotyping embryonic stem cells and mice exhibiting XbaI restriction sites and expected band sizes from a 5′ hybridizing probe on WT, targeted, and knockout alleles. (C) Southern screens generated from mouse tissue biopsy samples showing WT Mki67 bands at 14.0 kb and exon 4–deleted Mki67 bands at 3.7 kb. (D) In silico translation by ExPASy of mutant Ki67 protein sequence in comparison to WT Ki67 protein sequence. Red, initiating amino acid; blue, where open reading frame is read; -, where stop codon is read. (E) RT-qPCR analysis of the relative abundance of the sequences across the exon 3–4 junction in the Mki67 loci in liver samples from WT (n = 3) and Mki67^−/− mice (n = 3). (F) Western blot analysis of Ki67 protein expression in WT (n = 3) and Mki67^−/− (n = 3) splenocytes stimulated with 20 ng/ml LPS for 48 h. The expression of GAPDH was used as control. (G) Flow cytometry plot showing Ki67 protein expression in WT and Mki67^−/− splenocytes stimulated with 20 ng/ml LPS for 48 h. Data are representative of three mice from each group. (H) Histology showing H&E staining of skin, lung, liver, small intestine, BM, thymus, and spleen sections from WT and Mki67^−/− mice. Scale bars represent 20 μm. Data are representative of three mice from each group. Statistical significance between conditions in E was assessed by two-tailed unpaired t test with P value shown. Source data are available for this figure: SourceData FS1.