Figure 3

Reduction of Ih currents in entorhinal cortex layer II neurons of X11^-/-/X11L^-/-mice. ( A, B ) Representative electrophysiological recordings of Ih currents from EC layer II neurons in X11^+/+/X11L^+/+( A ) and X11^-/-/X11L^-/-( B ) mice. (C, D) Tail currents of X11^+/+/X11L^+/+ (C) and X11^-/-/X11L^-/- (D) mice. (E) Quantitative data of the voltage, whereby the current is half-activated (V1/2). Statistical analysis was performed using one-way analysis of variance followed by Tukey’s multiple comparison test (mean ± SEM (mV); five slices from two X11^+/+/X11L^+/+ mice, n = 10; four slices from two X11^-/-/X11L^+/+, X11^+/+/X11L^-/-, and X11^-/-/X11L^-/- mice, n = 9). (F) Quantitative data of series resistance. Statistical analysis was performed as described (mean ± SEM (MΩ); five slices from two X11^+/+/X11L^+/+ mice, n = 10; four slices from two X11^-/-/X11L^+/+, X11^+/+/X11L^-/-, and X11^-/-/X11L^-/- mice, n = 9). (G) Summary of the current density of the tail currents. Statistical analysis was performed as described (mean ± SEM (pA/pF); five slices from two X11^+/+/X11L^+/+ mice, n = 10; four slices from two X11^-/-/X11L^+/+, X11^+/+/X11L^-/-, and X11^-/-/X11L^-/- mice, n = 9; **p < 0.01). (H) Representative images of the HCN1-X11-X11L complex in EC layer II neurons of 13-week-old X11^+/+/X11L^+/+ mice (a, merge; b HCN1 (green); c, X11L (red); d, X11(blue)). Scale bar, 20 μm. (I−K) Co-immunoprecipitation of HCN1-X11s complexes from the 13-week-old X11^+/+/X11L^+/+ murine cortex. HCN1^-/- (I), X11^-/- (J), or X11L^-/- (K) mice were used as controls. Brain lysates were immunoprecipitated with anti-HCN1 (I), anti-X11 (J), and anti-X11L/Mint2 (K) antibodies. Immunocomplexes were detected by immunoblotting.