Fig. 6.

Generation of iRhom2^W538S/W538S mice. a The W538S mutation was inserted into exon 14 of the murine iRhom2 gene (RHBDF2) by CRISPR/Cas9. Genomic DNA (gDNA) and cDNA derived from generated mutant mice were analysed by PCR with primers specific for the wt or the W538S sequence. The possibility that introduced mutations accidentally altered the iRhom2 mRNA splicing was excluded by the fact that analysed cDNA of wt and W538S produced PCR products of the same size. As a negative control, H₂O was used instead of mRNA. To exclude primer binding to residual genomic DNA, a second negative control was also used. Here, the reverse transcriptase (RT) was omitted from the reaction. b mRNA expression of the ADAM17, iRhom1 and iRhom2 in liver, lung and spleen from mutant mice was measured by qPCR. n = 6