Figure 1: Usp16 is required for mouse embryonic development.
(a) Schematic representation of the strategies used to generate Usp16 knockout mice. Partial regions of Usp16 locus (from exons 2 to 7) are shown. Exons are shown as filled boxes and LoxP sites as filled triangles. PCR primers used for genotyping are shown as arrows. Position of cysteine 205, an amino acid essential for Usp16 deubiquitination activity, is indicated. (b) Identification of correctly targeted mouse embryonic stem cells. Top panels, phase contrast of the morphology of correctly targeted Usp16 ESCs as compared with wild-type V6.5 ESCs. Bottom panel, images of genotyping results of ESCs as shown in the top panel. PCR reactions were performed using primers as indicated in panel A. Scale bar, 50 μm. (c) Usp16 is required for mouse viability. Images of PCR genotyping of offspring from Usp16+/− mice intercrossing. The number and ratio of wild type and heterozygote adult mice genotyped were shown (the number in the parenthesis indicates litters examined). (d) Usp16 knockout results in early embryonic lethal. Image and PCR genotyping of E7.5 embryos from Usp16+/− mice intercrossing showing partially re-absorption of Usp16−/− embryos. The number and ratio of embryos genotyped were shown (the number in the parenthesis indicates litters examined). (e) Usp16−/− blastocysts are viable. PCR genotyping of blastocysts from Usp16+/− intercrossing. The number and ratio of blastocysts genotyped were shown under the PCR image (the number in the parenthesis indicates female mice killed). Original scans for gels are shown in Supplementary Fig. 7.
