FIG. 1.

Generation of Ube1L knockout mice. (A) Schematics of Ube1L knockout strategy. C on exon 14 indicates active site cysteine of UBE1L. (B) PCR genotyping of mouse tail DNA. (C) Northern blot analysis of Ube1L mRNA. A mixture of cells from spleen and thymus from mice of each genotype was cultured in vitro in the absence or presence of 100 U/ml of IFN-β for 12 h. The mRNA for the Ube1L gene was detected by Northern blotting. (D) Western blot analysis of protein ISGylation upon LPS challenge in mice. Eighteen hours after LPS injection, livers, lungs, and spleens were harvested, and protein ISGylation was detected by Western blotting using anti-mouse ISG15 polyclonal antibody. WT, wild type; HZ, heterozygous; KO, knockout.