Extended Data Fig. 10: Sex effect in MIA and three-hour dose response in maternal serum after MIA.
From: Prenatal immune stress blunts microglia reactivity, impairing neurocircuitry
a–b, Quantification of secreted IL-6 and TNFα from LPS-stimulated microglia in vitro at adulthood from the Str (a) and frontal Ctx (b) from Fig. 2a-b indicating the sex [male (blue) and female (pink)] of each offspring. n are independent culture experiments. Data are shown as mean ± s.e.m. with each experiment normalized to CON. Adult: nAll groups = 4 cultures each with 2 mice pooled from 3 litters for both CON and MIA. c–d, Baseline gene expression analysed by qPCR for Tnfα (c) and Aif1 (d) from bulk frontal Ctx and whole Str from CON (grey) and MIA (red) offspring indicating the sex [male (blue) and female (pink)] of each animal. n are individual animals. Data are shown as median ± IQR range with 1.5*IQR whiskers and analysed by a linear mixed-effect modelling and three-way ANOVA. Tnfα: nCON:CTX:female = 6 mice from 4 litters, nCON:CTX:male = 9 mice from 7 litters, nCON:STR:female = 6 mice from 4 litters, nCON:STR:male = 8 mice from 6 litters, nMIA:CTX:female = 5 mice from 4 litters, nMIA:CTX:male = 15 mice from 9 litters, nMIA:STR:female = 5 mice from 4 litters, nMIA:STR:male = 15 mice from 9 litters. Group: F(1, 65) = 3.78, p = 0.057, Region: F(1, 65) = 0.042, p = 0.84, Sex: F(1, 65) = 0.43, p = 0.51. Aif1: nCON:CTX:female = 6 mice from 4 litters, nCON:CTX:male = 2 mice from 2 litters, nCON:STR:female = 6 mice from 4 litters, nCON:STR:male = 2 mice from 2 litters, nMIA:CTX:female = 5 mice from 4 litters, nMIA:CTX:male = 3 mice from 2 litters, nMIA:STR:female = 5 mice from 4 litters, nMIA:STR:male = 3 mice from 2 litters. Group: F(1, 28) = 4.2, p=0.050, Region: F(1, 28) = 4.5, p=0.043, Sex: F(1, 28) = 1.5, p = 0.23. e, IL-6 protein expression in maternal serum 3 h after PIC injection. n are individual animals. Data are shown as mean ± s.e.m and analysed using a two-way ANOVA with Tukey posthoc test. *p < 0.05. ndose:0 = 81 litters, ndose:5 = 8 litters, ndose:10= 62 litters, ndose:20 = 28 litters. Group: F(3,162)=45.17, p < 2e-16, 0 v 5: t(162) = 1.7, p = 0.32, 0 v 10: t(162) = 9.8, p < 0.001, 0 v 20: t(162) = 9.4, p < 0.001, 10 v 20: t(162) = 1.7, p = 0.30. Offspring from dams with a serum response below 1,000 pg ml−1 were excluded from subsequent experiments. f. The gating strategy for FACS-isolated microglia used in Figs. 1j,k, 3 and 4a–j and Extended Data Figs. 2e, 5, 6 and 8
