Extended Data Fig. 1: Induction of apoE isoforms in microglia/CAMs in iE3/CreER/+ and iE4/CreER/+ mice. | Nature Immunology

Extended Data Fig. 1: Induction of apoE isoforms in microglia/CAMs in iE3/CreER/+ and iE4/CreER/+ mice.

From: Cell-autonomous effects of APOE4 in restricting microglial response in brain homeostasis and Alzheimer’s disease

Extended Data Fig. 1

a, b, Upon TAM administration, GFP fluorescence which represents apoE distribution was exclusively detected in the Iba1+ microglia (a, Scale bar, 40 µm) and TMEM119+ microglia (b, Scale bar, 20 µm) in the iE3/CreER/+ and iE4/CreER/+ mice but not in control mice. The percentage of GFP+ cells in Iba1+microglia or TMEM119+ microglia in experimental mice (iE3/CreER/+: Ctrl: n = 5, TAM-induced: n = 5; iE4/CreER/+: Ctrl: n = 5, TAM-induced: n = 5) was quantified. **, P < 0.0001. Two-tailed t-test. c, Brain tissues from TAM-induced mice were co-immunostained for GFP which represents apoE distribution (green), and astrocyte-specific (anti-GFAP; pink) or neuron-specific (anti-NeuN; red) markers. Experiments were repeated two times with similar results. Scale bar, 20 µm. d, The GFP fluorescence was colocalized with apoE signal (Red) detected by apoE antibody. Scale bar, 20 μm. e, The percentage of apoE+ cells in GFP+ microglia in iE3/CreER/+ (Ctrl: n = 5, TAM-induced: n = 8) and iE4/CreER/+ (Ctrl: n = 5, TAM-induced: n = 8) mice was quantified. **, P < 0.0001. f, ApoE levels in the cortex of the iE3/CreER/+ and iE4/CreER/+ mice at 6 months of age upon vehicle or TAM administration (iE3/CreER/+: Ctrl: n = 7, TAM-induced: n = 5; iE4/CreER/+: Ctrl: n = 7, TAM-induced: n = 5) were analyzed by ELISA. **, P < 0.0001. g, ApoE levels in the cortex of the iE3/CreER/+ (Ctrl: n = 8, TAM-induced: n = 8) and iE4/CreER/+ (Ctrl: n = 9, TAM-induced: n = 7) mice were examined by Western blotting and quantified. **, P < 0.0001. h, ApoE levels in the plasma of the iE3/CreER/+ (Ctrl: n = 5, TAM-induced: n = 5) and iE4/CreER/+ (Ctrl: n = 5, TAM-induced: n = 3) mice upon vehicle or TAM administration were analyzed by ELISA. Plasma from APOE3-TR and APOE4-TR mice (n = 4/group) was included as positive controls. **, P < 0.0001, one-way ANOVA. a, b, g-h, Data represent mean ± s.e.m. N.S., not significant. e-g, two-way ANOVA with a Tukey’s post-hoc test.

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