Fig. 4
Characterization of whole-body RelA-/- and RelACKO/CKO MEF cells. a Mating of a RelA+/- male and female mice produced RelA+/+, RelA+/- and RelA-/- embryos that were isolated on E12. White arrow indicates the liver, which was observed macroscopically in RelA+/+ and RelA+/- embryos only, whereas a large hematoma was observed in abdomens of RelA-/- embryos. b Immunohistochemistry for cleaved-caspase 3 and cleaved-PARP demonstrated paucity of immunostaining in WT embryos for but very robust staining for the enzymes in KO embryos indicative of massive liver apoptosis. c PCR was performed on DNA isolated from the embryos. Presence of the 7–8 product (425 bp) indicates presence of a WT allele whereas presence of the 5–8Δ product (335 bp) indicates a RelA mutant allele. d QRT-PCR on RNA isolated from MEF cells provides evidence that RelA transcripts decreased with decrease in RelA WT allele. e Western blot of proteins extracted from the same MEF cells probed with anti-RelA Ab (top). β actin staining was used as loading control (bottom). There was no shift in the RelA band suggesting absence of any truncated form of RelA. These data suggest decrease in the RelA protein with decreasing dose of WT RelA DNA. f RelACKO/CKO MEF cells were transduced with lentiviral-Cre at multiplicity of infection (MOI) of 1 and 2. Induction of Cre led to a substantial decrease in RelA with increasing MOI