Figure 3.
Validation and characterization of NEMOLPC-KO/3DRLPC-KO mice. (a) qRT-PCR analysis was performed using total liver RNA (n=5 mice per genotype, expression normalized to Tbp). (b) PCR genotyping on genomic DNA isolated from total liver or tail tissue. F, floxed allele; D, deleted allele. (c) Graph showing deletion efficiency of TNFR1 and TRAIL-R measured using quantitative PCR analysis of genomic DNA from total liver tissue or isolated hepatocytes. (d and e) Immunoblot analysis of liver lysates with the indicated antibodies (d) and immunostainings of liver sections with antibodies recognizing cleaved caspase-3 (e) from NEMOLPC-KO and NEMO-3DRLPC-KO mice 5 h after TNF or PBS administration. (f) FACS analysis of surface Fas expression in hepatocytes isolated from mice with the indicated genotypes. (g) Immunoblot analysis of hepatocyte protein extracts from NEMOLPC-KO/3DRLPC-KO and control showing deletion of NEMO in hepatocytes isolated from mice