Fig. 5: PV interneurons show significantly reduced perisomatic innervation in Tg(Nkx2.1-Cre);Tsc1^flox/flox and Tg(Nkx2.1-Cre);Tsc1^flox/+ mice at EP34.

a A PV cell (green) among NeuN immunostained neurons (blue) in cortical organotypic cultures from a Tsc1^Ctrl mouse at EP34. b, c PV cells from Tg(Nkx2.1-Cre);Tsc1^flox/+ mice (b1–b3) or Tg(Nkx2.1-Cre);Tsc1^flox/flox mice (c1–c3) show decreased bouton density (d) (one-way ANOVA, *p = 0.0157; Holm–Sidak’s multiple comparisons: Tsc1^Ctrl vs Tg(Nkx2.1-Cre);Tsc1^flox/+ *p = 0,0214; Tsc1^Ctrl vs Tg(Nkx2.1-Cre); Tsc1^flox/flox *p = 0.0214). Local branching (e) (one-way ANOVA, **p = 0.0040 (Radius 7), *p = 0.0127 (Radius 8), **p = 0.0011 (Radius 9); Holm–Sidak’s multiple comparisons: (Radius 7) Tsc1^Ctrl vs Tg(Nkx2.1-Cre);Tsc1^flox/+ **p = 0.0032; Tsc1^Ctrl vs Tg(Nkx2.1-Cre); Tsc1^flox/flox *p = 0.0117; (Radius 8) Tsc1^Ctrl vs Tg(Nkx2.1-Cre); Tsc1^flox/+*p = 0.0126; Tsc1^Ctrl vs Tg(Nkx2.1-Cre); Tsc1^flox/flox *p = 0.0149; (Radius 9) Tsc1^Ctrl vs Tg(Nkx2.1-Cre); Tsc1^flox/+ ***p = 0.0008; Tsc1^Ctrl vs Tg(Nkx2.1-Cre);Tsc1^flox/flox **p = 0.0034). Number of mice: Tsc1^Ctrl n = 6, Tg(Nkx2.1-Cre);Tsc1^flx/+ n = 6, Tg(Nkx2.1-Cre); Tsc1^flox/flox n = 6. f Percentage of innervation is also significantly lower for PV cells from Tg(Nkx2.1-Cre);Tsc1^flox/+ and Tg(Nkx2.1-Cre); Tsc1^flox/flox mice (one-way ANOVA, *p = 0.0212, Holm–Sidak’s multiple comparisons: Tsc1^Ctrl vs Tg(Nkx2.1-Cre);Tsc1^flox/+ *p = 0.0257; Tsc1^Ctrl vs Tg(Nkx2.1-Cre); Tsc1^flox/flox *p = 0.0179). Number of PV cells: Tsc1^Ctrl n = 6, Tg(Nkx2.1-Cre);Tsc1^flox/+ n = 8, Tg(Nkx2.1-Cre);Tsc1^flox/flox n = 6. Arrowheads indicate boutons. Scale bars: a1–c1, 20 µm; a2–c2, 10 µm, a3–c3, 3 µm. Data represent mean ± SEM. Source data are provided as a Source Data file.