Site-directed mutagenesis of the CD11c^1000bp promoter sequence. (A) The original CD11c^1000bp promoter sequence (top row) was modified by site-directed mutagenesis. The predicted TFBSs are represented by boxes, and the mutated bases are highlighted in red (bottom row). Because of the repetitive sequence structure in the BRNF binding region, additional mutations had to be introduced. (B) Flow cytometric analysis of promoter activity in DCs generated in vitro. At day 1 or 2 of the granulocyte-macrophage colony-stimulating factor culture, bone marrow cells were transduced with lentiviral vectors (CD11c^1000bp-GFP and CD11c^{1000bp-mutated}-GFP) at a multiplicity of infection of 1, and GFP expression was analyzed at day 7 of the culture. The control was only treated with polybrene. (C) Results from 2 independently performed experiments are normalized (GFP expression of the CD11c^1000bp construct set to 100%). (D) In vivo comparison of the CD11c^1000bp and the CD11c^{1000bp-mutated} promoters by flow cytometric analysis of splenocytes from bone marrow chimeras, which were generated and analyzed analogously to Figure 3B. Data are representative of 2 independently performed experiments with 3 mice per group.