Figure 6. Whole Mount in situ Hybridization of Targets Down-regulated by Lmx1b.

WMISH for Q-PCR validated down-regulated genes at 12.5 dpc, cut sections performed as illustrated by cartoon in Fig. 4. Rfx4 is expressed dorsally (A) and ventrally (A′, white arrowhead; A”, cut section, white arrowhead) WT limb. In the Lmx1b KO limb, Rfx4 is expressed dorsally (B) and ventrally (B′). A cut section shows duplicated expression (B”, white arrowhead). C and C′ show dorsal and ventral Aff3 expression in WT limb, respectively. C” displays enhanced ventral Aff3 expression, magnification area denoted by the white box demonstrates absence of Aff3 in the dorsal subectodermal mesoderm (red line). D and D′ show dorsal and ventral expression of Aff3 in the Lmx1b KO limb, respectively. D” magnified in d″, Aff3 is enhanced both dorsally and ventrally limb.