Figure 1. Expression profiling of wild-type and p100^−/− MEFs.

(A) Western blot analysis of cytoplasmic and nuclear protein extracts (20 µg/sample) were analyzed for the presence of NF-κB family members p100/p52, RelB, and RelA in wild-type (+/+) and in p100^−/− (−/−) MEFs. As cytoplasmic loading control S6 ribosomal protein and as nuclear loading control RNA Pol II was assayed. (B) Increased κB DNA-binding activity in nuclear extracts from p100^−/− (−/−) compared to wild-type MEFs (+/+). Five µg protein extract per cell line were incubated with a radioactively labeled Igκ oligo and analyzed by EMSA. Supershift analysis was performed using pre-immune serum (p.i.), anti-RelA (α-RelA), anti-RelB (α-RelB), and anti-p52 antibodies (α-p52). Super-shifted RelB and p52 complexes are indicated by arrow and arrowhead, respectively. (C) Heatmap displaying fold change values observed in p100^−/− vs. wild-type cells. The color code indicates the fold change values between +6 fold up- (red) and −20 fold downregulation (green). Each horizontal line on the heatmap corresponds to one gene. Genes labeled with blue boxes on the left were verified by qRT-PCR. Gene symbols and abbreviations of GO terms are displayed on the right. CA, Cytokine activity; GPCRB, G-protein-coupled receptor binding; IGFB, insulin-like growth factor binding; ER, extracellular region; ESO, extracellular structure organization and biogenesis; D/M, developmental process; B/ND, forebrain development and nervous system development; CG/S, regulation of cell size; IR/RES, immune response and response to external stimulus; M/L/T, locomotory behavior. (D) Significantly regulated Gene Ontology terms with respective gene numbers.