Extended Data Fig. 6: Fezf2 deletion causes a spatial redistribution of mid-layer interneuron subtypes.
From: Pyramidal neurons proportionately alter cortical interneuron subtypes
a, b, RNAscope ISH confirms an upward laminar shift of the total PVALB (a) and SST (b) interneuron populations in the SSp of P28 Fezf2 KO mice. Scale bar, 100 µm. Bar plots (right) quantify cell proportion per layer (mean ± SD). Sample sizes are n = mice (ROIs). control: n = 5 (10), age P26-28, n = 3932 PVALB, n = 2280 SST. Fezf2 KO: n = 3 (5), age P27-28 n = 2377 PVALB, n = 1677 SST. Statistical significance was determined using a LMM with animal ID as a random effect. n.s., not significant, P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. c, Bar plots showing the composition of each PVALB and SST interneuron subtypes in different cortical layers of control and Fezf2 KO brains in the SSp region, based on MERFISH dataset. control: n = 2 (3), Fezf2 KO: n = 3 (3), age: P14-30. d, Representative RNAscope ISH images of SST–Calb2 interneurons in control and Fezf2 KO SSp cortex, which show no obvious laminar shift. Signal is visualized by a Calb2–Vip subtraction. Note: this strategy also labels L6 SST–Chodl interneurons (n = 3 control, n = 3 KO; P28). e, RNAscope ISH images of Hpse mRNA in SSp of control and Fezf2 KO brains, showing a denser and broader accumulation of SST-Hpse interneurons near the L4/L5 boundary in the Fezf2 KO (arrow). Note: L5 PNs express low levels of Hpse (n = 4 control, n = 5 KO; P27-P31). f, Representative images of viral genetically labeled SST–Hpse interneurons in control and Fezf2 mutants at P47 (n = 1 KO; cohort size was ethically limited due to high post-operative mortality in the HpseCre;Fezf2lacZ/lacZ background). For d-f, Scale bar: 100 µm. Exact sample genotypes, statistical model details, and P values are provided in Supplementary Tables 2 and 3.
