Fig. 6

Blocking maternal IFN-γ signaling mitigates the effects of MIA on offspring. (a) Schematic diagram of the experimental design. Pregnant dams were pretreated with IFNGR-blocking antibodies (anti-IFNGR) or IgG before injecting Poly (I: C) or PBS on E12.5. (b) Statistical graphs of ASD-like behaviors in offspring, including three-chamber social test, grooming test, and marble burying test (n = 9–11 from 4–5 litters). Two-way ANOVA tests (for the three-chamber social test) and Unpaired T tests (for grooming and marble burying tests) were applied. *** P < 0.001, ** P < 0.01, * P < 0.05, ### P < 0.001, # P < 0.05 and ns P > 0.05. (c) Representative MRI images and quantification of LVs areas in offspring (n = 7–8 from 4–5 litters). Scale bar, 5 mm. One-way ANOVA tests were applied. ** P < 0.01. (d) Quantification of CSF production rate in offspring (n = 3–4 from 3 litters). Unpaired T tests were applied. ** P < 0.01. (e) Quantification of the CSF fluorescence intensity in offspring from the B-CSFB assessment (n = 5 from 3–4 litters). Unpaired T tests were applied. * P < 0.05. (f and i) Representative IF staining images (f) and quantification (i) pNKCC1 fluorescence intensity in the Chp of offspring (n = 5–6 from 3–4 litters). Scale bar, 100 μm. One-way ANOVA tests were applied. * P < 0.05. (g and j) Representative IF staining images (g) and quantification (j) of Iba1⁺ macrophage in the Chp of offspring (n = 4–6 from 3–4 litters). Scale bar, 100 μm. One-way ANOVA tests were applied. ** P < 0.01. (h and k) Representative IF staining images (h) and quantification (h) of Occludin and β-Catenin fluorescence intensity in the Chp of offspring (n = 4–5 from 3–4 litters). Scale bar, 100 μm. One-way ANOVA tests were applied. ** P < 0.01 and *** P < 0.001. (l) Representative images of fluorescent microsphere trajectories. Scale bar, 100 μm. Quantification of fluorescent microsphere movement speed in the LVs of offspring (n = 6, from 3 litters per group). Unpaired T tests were applied. ** P < 0.01. (m and o) Representative IF staining images (m) and quantification (o) of ciliary tufts in the LVs of offspring (n = 4–5 from 3 litters). Scale bar, 100 μm. One-way ANOVA tests were applied. *** P < 0.001, ** P < 0.01. (n-r) Representative IF staining images of ECs in the lateral ventricles of offspring (n). Scale bar, 100 μm. Quantification of the density of Foxj1⁺Sox2⁺ ECs (p) and Foxj1⁻Sox2⁺ neural progenitor cells (q) and the ratio of the two (r), (n = 5–6 from 3–4 litters). Unpaired T tests were applied. ** P < 0.01 and * P < 0.05