Bone Marrow-Specific Knock-In of a Non-Activatable Ikkα Kinase Mutant Influences Haematopoiesis but Not Atherosclerosis in Apoe-Deficient Mice
Figure 7
IkkαAA/AA knock-in does not influence macrophage lipid uptake.
(A) Flow cytometric analysis of Dil-oxLDL uptake by BM-derived macrophages. Cells were incubated without or with Dil-oxLDL (1 µg/ml or 10 µg/ml) for 3 h or 24 h, as indicated. To test for actin-dependent uptake, additional controls were simultaneously treated with cytochalasin D (cytD), as indicated. Representative flow cytometric histograms are shown. Graphs represent the mean ± SEM (n = 3); 2-way ANOVA with Bonferroni post-test. (B–D) Intracellular lipid depositions in aortic root lesions were stained with Nile Red and co-stained with Mac2 in order to quantify lipid-laden macrophages. (B) Nile Red staining was quantified relative to the plaque area (left graph), and Nile Red+ cells were quantified as percentage of total plaque cells (right graph). (C) Lipid uptake by macrophages was quantified as Nile Red+Mac2+ area as % of Mac2+ area (left graph), and as Nile Red+Mac2+ cells as % of Mac2+-cells. (D) Shown are representative pictures from Nile Red staining (red fluorescence; left image). The middle and right image demonstrate Image J analyses. Displayed are the plaque area (thin red line), macrophage area (green), plaque cell nuclei (blue), Nile Red+ area (yellow) and Mac2+ Nile Red+ area (white, lipid deposits in macrophages). Graphs represent the mean ± SEM (n = 5–6).