Extended Data Fig. 7: Astrocytes recDEGs knock-down alters astrocytes states in vitro. | Nature Neuroscience

Extended Data Fig. 7: Astrocytes recDEGs knock-down alters astrocytes states in vitro.

From: Protective exercise responses in the dentate gyrus of Alzheimer’s disease mouse model revealed with single-nucleus RNA-sequencing

Extended Data Fig. 7: Astrocytes recDEGs knock-down alters astrocytes states in vitro.

Primary cortical mixed glia cultures were transduced with LV-shRNA for five days in standard growth media (a-d) or treated in the last 24 h with a reactive astrocyte activating cocktail of cytokines (e-h) or amyloid-β 42 peptide (i-l). Confirmation of the gene knock-down by QPCR (a, e, i), and gene expression of reactive astrocyte markers and markers of our subcluster 0 and 1 in response to Nme7 (b, f, j), St7 (c, g, k), and Thra (d, h, l) knock-down. n = 3 for shCtrl in e-l, n = 4 for all other groups. Two-way ANOVA followed by Fisher’s LSD. *p < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s., not significant. Data represent the mean ± s.e.m. of biologically independent samples. a: two-way ANOVA, KD and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl ****P < 0.0001; b: two-way ANOVA, KD and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl Gfap, Cdh20, Rorb, and Cdh4 ****P < .0001, C3 n.s. P = 0.3171, Serpina3n ***P = 0.0009, Hspb1 **P = 0.004, Csmd1 **P = 0.0068; c: two-way ANOVA, KD n.s. P = 0.2011, KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl Gfap, C3, Serpina3n, Cdh20, and Cdh4 ****P < 0.0001, Hspb1 *P = 0.0277, Rorb n.s. P = 0.1921, Csmd1 n.s. P = 0.9147; d: two-way ANOVA, KD and KD x gene ****P < .0001, followed by Fisher’s LSD compared to shCtrl Gfap, Serpina3n, Rorb, Cdh4, and Csmd1****P < 0.0001, C3 ***P = 0.0001, Hspb1 n.s. P = 0.283, Cdh20 *P = 0.0174; e: two-way ANOVA, KD and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl ****P < .0001; f: two-way ANOVA, KD and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl Gfap, Serpina3n, Cdh20, and Rorb ****P < 0.0001, C3 n.s. P = 0.7271, Hspb1 n.s. P = 0.107, Cdh4 n.s. P = 0.0856, Csmd1 n.s. P = 0.1875; g: two-way ANOVA, KD and KD x gene ****P < .0001, followed by Fisher’s LSD compared to shCtrl Gfap **P = 0.0092, C3, Cdh20, Cdh4, and Csmd1 ****P < .000, Serpina3n **P = 0.0012, Hspb1 ***P = 0.001, Rorb n.s. P = 0.159; h: two-way ANOVA, KD and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl Gfap, Serpina3n, Cdh20, Rorb, and Cdh4****P < 0.0001, C3 n.s. P = 0.8352, Hspb1 n.s. P = 0.9755, Csmd1 n.s. P = 0.1847; i: two-way ANOVA, KD and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl ****P < 0.0001; j: two-way ANOVA, KD and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl Gfap ***P = 0.0002, C3 n.s. P = 0.2851, Serpina3n **P = 0.003, Hspb1 n.s. P = 0.5116, Cdh20, Rorb, and Cdh4 ****P < 0.0001, Csmd1 **P = 0.0093; k: two-way ANOVA, KD n.s. P = 0.7698 and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl Gfap, ***P = 0.0001, C3, Cdh20, and Cdh4 ****P < 0.0001, Serpina3n n.s. P = 0.137, Hspb1 n.s. P = 0.0896, Rorb n.s. P = 0.6144, Csmd1 n.s. P = 0.6805. l: two-way ANOVA, KD n.s. P = 0.4382 and KD x gene ****P < 0.0001, followed by Fisher’s LSD compared to shCtrl Gfap, Serpina3n, Hspb1, Cdh20, Rorb, and Cdh4 ****P < 0.0001, C3 n.s. P = 0.9321, Csmd1 * P = 0.0236.

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