Fig. 4. Generation and verification of mice expressing a deletion of Smoothened in the myeloid cell lineage (LysMCre/Smo^KO mice).

(A) A mouse model expressing a myeloid cell-specific deletion of Smoothened (LysMCre/Smo^KO) was generated using transgenic mice bearing loxP sites flanking exon 1 of the Smo gene (Smo loxP) and mice expressing a Cre recombinase transgene from the Lysozyme M locus (LysMCre). (B) Genotyping was based on polymerase chain reaction primers specific for Cre, mutant Smo and wild type Smo (Smo WT). Shown are the fragments amplified from LysMCre/Smo^KO, WT control and LysMCre/Smo^het (heterozygous for WT and mutant Smo) mice. (C) Representative flow cytometric contour plots of the gating scheme for CD11b^highF4/80^highLy6C^highLy6G^low macrophages in peripheral blood collected from control recipients transplanted with control donor (cont^cont) or LysMCre/Smo^KO donor (cont^SmoKO) bone marrow cells. (D) RNA was extracted from FACS sorted macrophages and analyzed for expression of smoothened. Shown is the fold change relative to the cont^cont group.