ALT Figure 4 Manipulation of Hh signaling in A. sagrei embryos and primary cells. (A) Schematic of in ovo treatment of lizard embryos with SAG. Each egg was collected within 24 h of being laid and placed on top of a SAG or DMSO solution. These solutions were replaced by water after 24 h. (B) Embryos were collected from SAG- or DMSO-treated eggs after 14 days of incubation. Blue arrows highlight the location of extra digits. FL, forelimbs; HL, hindlimbs. Scale bar: 500 µm. (C) Schematic of primary lizard embryonic fibroblast cell (LEF) SAG treatment. Cells from either the torso (region between dashed yellow lines) or limb buds (outlined in blue) from a single embryo were grown in culture before being treated with SAG or DMSO