Bone Marrow-Specific Knock-In of a Non-Activatable Ikkα Kinase Mutant Influences Haematopoiesis but Not Atherosclerosis in Apoe-Deficient Mice
Figure 6
IkkαAA/AA knock-in does not enhance or prolong NF-κB p65 activity, or majorly influence cytokine expression in Apoe−/− macrophages in vitro.
(A) BM-derived macrophages from IkkαAA/AAApoe−/− and Ikkα+/+Apoe−/− mice were stimulated in vitro with 10 ng/ml Tnf-α, 50 µg/ml of mildly oxidized LDL or 100 ng/ml LPS for the indicated time. Activation of p65 was quantified in nuclear extracts using a TransAm p65 assay. Graphs represent the mean ± SEM (n = 2); 2-way ANOVA with Bonferroni post-test, ***P<0.001. (B) BM-derived macrophages from IkkαAA/AAApoe−/− and Ikkα+/+Apoe−/− mice were stimulated in vitro for 24 h with 10 ng/ml Tnf-α or 50 µg/ml heavily oxidized LDL. Cytokine concentrations in the supernatants are displayed for Tnf-α, Il-6 and Mcp1. Graphs represent mean ± SEM (n = 9 from 3 independent experiments); 2-way ANOVA with Bonferroni post-test, ***P<0.001. (C) Concentrations of Tnf-α, Mcp1 and Il-6 in serum of Apoe−/− mice transplanted with IkkαAA/AAApoe−/− or Ikkα+/+Apoe−/− BM and receiving a high-fat diet for 8 weeks. Graphs represent the mean ± SEM (n = 7–9).