Fig. 8: Blocking AR rescues the enhanced cortical neurogenesis and autistic behavior in male offspring caused by MeCP2 duplication.

A Schematic of the experimental design and flutamide (FT) treatment timeline. WT and MeCP2^Tg1 male embryos were injected intracerebrally at E14.5 with vehicle (10% DMSO in saline) or FT (0.6 μg), and analyzed at E18.5. B, C Immunofluorescence of MEF2C⁺ cells in the cerebral cortex of vehicle-treated WT, vehicle- and FT-treated MeCP2^Tg1 mice. B Representative images and distribution of MEF2C⁺ cells across 10 cortical bins (N = 6 males from 6 dams per group). (C) Quantification of MEF2C⁺ cell number per 100 μm bin (N = 8 males from 6 dams per group). Scale bars, 50 μm. Red and blue labels indicate comparisons between WT + Veh. vs MeCP2^Tg1+ Veh. and MeCP2^Tg1+ Veh. vs MeCP2^Tg1 + FT, respectively. D Immunostaining and quantification of SATB2⁺ upper-layer neurons in the cortex (N = 7 males from 6 mothers per group). Scale bar, 50 μm. E SOX2⁺ progenitor cells were analyzed in vehicle- and FT-treated MeCP2^Tg1 cortices (N = 8 males per group). Representative images and high-magnification views of boxed areas are shown. Quantification was based on cell counts per 150 μm bin. F, G Three-chamber social interaction (F) and social novelty (G) tests. Diagrams are shown on the left, and quantifications are shown on the right. N = 18 males from 10 dams per group. H Marble burying index, calculated as the proportion of buried marbles (N = 21 males from 10 dams per group). I–K Open field test: representative locomotor heatmaps (I), total distance traveled (J), and center/total distance ratio (K), N = 20 males from 10 dams per group. L, M Elevated plus maze test: representative locomotion tracks (L) and quantification of open arm distance (M) (N = 20 males from 10 dams per group). Data are presented as mean ± SEM. Box plots show the median (center line), 25th–75th percentiles (box limits), and whiskers indicate the minimum and maximum values. Exact P-values are indicated. Statistical analyses were performed using two-way ANOVA test followed by Sidak post-hoc test for (B), and one-way ANOVA followed by Tukey post-hoc test for (C–H, J, K and M). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.