(A) HSCs were isolated from Smo-flox mice. AdGFP or AdCre (MOI 25) were added on culture day 4, and cells were harvested 3 days later. Effects on gene expression were assessed by qRT-PCR. Results are normalized to day 0 HSCs. *P < 0.05. Inset shows representative Western blot for expression of Cre and tubulin. (B) Oligonucleotides for the floxed SMO allele (refs. 56, 63; Supplemental Table 3) were used to distinguish the unrearranged allele (Smo^c; 1.7 Kb) from the Cre-recombined null allele (Smoⁿ; 0.5 Kb). Genomic DNA was isolated from livers of DTG mice treated either with vehicle (VEH, n = 3) or TMX (n = 4) from day 4–10 after BDL, and Smoⁿ and Smo^c amplicons were amplified. Quantification of the amplicons was performed by densitometry to assess the efficiency of Cre-mediated recombination. Results are expressed as Smoⁿ/Smoⁿ + Smo^c x 100, shown below each lane. (C) Total RNA was isolated from livers of vehicle- and TMX-treated DTG mice 14 days after sham surgery or BDL. SMO expression was assessed by qRT-PCR, and results are expressed as fold over sham plus vehicle control. *P < 0.05. (D) Representative immunohistochemistry for hepatic GLI2. GLI2⁺ cells in vehicle-treated and TMX-treated DTG mice were quantified in sham and BDL groups and represented as number of positive cells per field (n = 11/group; original magnification, ×20 [Sham]; ×63 [BDL]). *P < 0.05, **P < 0.01. (E) Total RNA was isolated from livers of vehicle- and TMX-treated DTG mice 14 days after BDL and analyzed by qRT-PCR for expression of Hh-regulated genes. Results are expressed as fold over vehicle-treated group. *P < 0.05.