Supplementary Figure 1: Hdac3 forebrain-specific conditional knockout from excitatory neurons.

(a) Coronal brain slices of 3-month control and Hdac3 cKO male mice were stained using Hoechst (blue) and anti-HDAC3 (green), and images captured of the hippocampus using tile scan of a confocal microscope. Scale bar, 200 μm. (b) Mean gray value quantification of HDAC3 immunofluorescence was compared between control and Hdac3 cKO mice in the CA1 and dentate gyrus (DG) of the hippocampus; two-tailed t test; CA1, t₍₈₎ = 12.88, **** P < 0.0001; DG, t₍₈₎ = 5.958, *** P = 0.0003; n = 4, 6 mice. (c) Coronal brain slices of 3-month control and Hdac3 cKO male mice were stained using Hoechst (blue), anti-HDAC3 (red), anti-GFAP (green), and anti-NeuN (white), and images captured of hippocampal CA1 using a confocal microscope. Scale bar, 20 μm; n = 3, 3 mice. (d) Coronal brain slices of 3-month control and Hdac3 cKO male mice were stained using Hoechst (blue), anti-HDAC3 (red), anti-Parvalbumin (green), and anti-NeuN (white), and images captured of hippocampal CA1 using a confocal microscope. Scale bar, 20 μm; n = 3, 3 mice. (e) Western blot analysis of HDAC3, HDAC2 and actin protein levels using hippocampal whole cell lysates from control and Hdac3 cKO mice. Full-length WB’s are presented in Supplementary Fig. 10b. (f) Mean gray value quantification of HDAC3 protein levels normalized to actin of western blot in c. Two-tailed t test; t₍₆₎ = 9.663, **** P < 0.0001; n = 4, 4 mice. (g) Western blot analysis of HDAC3 and actin protein levels using cortex whole cell lysates from control and Hdac3 cKO mice. Full-length WB’s are presented in Supplementary Fig. 10c. (h) Mean gray value quantification of HDAC3 protein levels normalized to actin of western blot in e. Two-tailed t test; t₍₆₎ = 3.460, * P = 0.0135; n = 4, 4 mice. (i), Western blot analysis of HDAC3 and actin protein levels using striatum whole cell lysates from control and Hdac3 cKO mice. Full-length WB’s are presented in Supplementary Fig. 10d. (j) Mean gray value quantification of HDAC3 protein levels normalized to actin of western blot in g. Two-tailed t test; t₍₆₎ = 0.9516, P = 0.3780; n = 4, 4 mice. All values are mean ± s.e.m.