Figure 6.

In vivo genome-wide expression analysis reveals that Lmx1b regulates genes involved in neuronal development, including En1 and En2 and acts as a repressor of Ahd2 expression. (A) Schematic representation of the experimental setup for sample preparation and sample comparison. (B) Among the 20 most regulated genes are Ahd2 (Aldh1a1, red), which shows an 85% increase, and En1 (red) and En2 (red) which are both down-regulated (En1, ~25% loss, En2, ~27% loss). (C) Venn diagram illustrating that nine genes regulated by Lmx1b are mdDA enriched. (D) Relative expression of nine genes that were mdDA enriched. Among the 9 genes En1 and Ahd2 were considered most interesting (red arrows). (E) Relative expression of genes regulated by both En1 and Lmx1b demonstrate that genes that are regulated by the loss of En1 (gray bars) are affected in the same manner in the Pitx3Cre/+ Lmx1b L/L E14.5 embryos (black bars). Wildtype was set at 1. (F) When looking at the relative expression of genes regulated by Pitx3 and Lmx1b the opposite effect is observed as with the En1 mutant. Genes up-regulated in the Pitx3 mutant are down-regulated in the Lmx1b depleted embryos and genes up-regulated in the Pitx3Cre driven Lmx1b mutant are down-regulated in embryos missing Pitx3. (G) Venn diagram showing that Lmx1b, En1 and Pitx3 are all involved in regulating En1, En2 and Cd9.