Hematopoietic-specific deletion of Ptch1. (A) Ptch1 Southern blot of BM cells from SclPtch1^fl/fl and SclPtch1^+/fl mice 8 weeks after tamoxifen. Unlike all other blots, the probe used for this blot distinguishes Ptch1 wild-type (+) and Ptch1^fl alleles. (B) Representative dot-plots of lineage-negative BM cells from SclPtch1-heterozygous (+/Δ) and SclPtch1-null (Δ/Δ) mice 8 weeks after tamoxifen. (C) PCR for efficiency of Ptch1^fl deletion in pre-BII cells FACS-isolated from a CD19Ptch1^Δ/Δ mouse. Deletion efficiency is at least 80%, as determined by the dose titration curve shown on the left. (D) Pre-B cell subsets in BM from Ptch1^fl/fl (fl/fl) and CD19Ptch1-null (Δ/Δ) mice. Mean plus SD calculated from 4 mice of each genotype. (E) Thymocyte cellularity of Ptch1^fl/fl (fl/fl) and LckPtch1-null (Δ/Δ) mice. Mean plus SD calculated from 4 mice of each genotype. (F) Representative dot-plots of T-cell subsets. The mean percentage shown for each subset was calculated from 4 mice of each genotype. (G) Quantitative RT-PCR for Gli1 mRNA expression in the mesenchymal cell line C3H treated with or without conditioned media containing Shh. Mean plus SD for 3 independent experiments is shown. (H) Quantitative RT-PCR for Gli1 mRNA expression in FACS-isolated BM fractions (pre-BI, pre-BII, LKS, and LK) and thymocyte fractions (DN and DP) from control (Ptch^fl/fl) and MxPtch1-null (Δ/Δ) mice. Results for B- and T-cell subsets are mean plus SD of 2 independent sorts, and LKS and LK results are from a single sort from a pool of 5 mice of each genotype.