The goal of reducing Kcnj6 gene dose in Ts65Dn mice was to reduce Kir3.2 protein levels to those observed in 2N animals. To determine if this goal was reached, protein levels in the hippocampi of individual mice were assessed using western blot. For side by side comparison, three experimental series were performed with the samples from the experimental groups loaded serially (Fig. 2). As expected, in Ts65Dn:Kcnj6+++ mice the levels of Kir3.2 were increased (F1, 6 = 3.04, p = 0.01), while the levels of Kir3.1 unchanged (F1, 9 = 0.59, p = 0.29) relative to 2N:Kcnj6++ mice (Fig. 2A). The increase in Kir3.2 levels approximated the increase in Kcnj6 dosage in Ts65Dn:Kcnj6+++ vs. 2N:Kcnj6++ mice. Removal of the extra copy of Kcnj6 reduced Kir3.2 levels and had no effect on Kir3.1. Side by side comparison of Ts65Dn:Kcnj6++− vs. 2N:Kcnj6++ samples showed no difference in the protein levels in these experimental groups (Kir3.1: F1,6 = 0.09, p = 0.46; Kir3.2: F1,9 = 0.01, p = 0.49) (Fig. 2B). Comparison of Ts65Dn:Kcnj6++− vs. Ts65Dn:Kcnj6+++ samples again showed a reduction of Kir3.2
