Journal: Molecular Psychiatry

Article Title: C/EBPβ is a key transcription factor for APOE and preferentially mediates ApoE4 expression in Alzheimer’s disease

doi: 10.1038/s41380-020-00956-4

Figure Lengend Snippet: Viral injection of ApoE3 or ApoE4 into C/EBPβ+/+; C/EBPβ+/−; 3xTg and 3xTg/C/EBPβ+/− mice (3 months old) for 3 months. A The hippocampal tissues were analyzed by immunoblotting (n = 2 mice per group, arrow: full-length AEP, arrowhead: activated AEP). B, C Overexpression of ApoE4 increased the activity of AEP and expression of Aβ. Each mouse is analyzed three times and the averaged values are plotted and subjected to statistical analyses (n = 3 mice/group, mean ± SEM, **P < 0.01, two-way ANOVA and Bonferroni’s post hoc test). D ApoE4 overexpression enhanced the early formation of amyloid plaques in wild-type or 3xTg mice, and downregulation of C/EBPβ decreased Aβ aggregates, co-staining with Aβ and THS, Scale bar: 100 μm. E Quantification from both 4G8 and THS-positive signals, n = 5, 2 mice/group, 2–3 slices/mouse (mean ± SEM, *P < 0.05, **P < 0.01 vs. group 1, two-way ANOVA and Bonferroni’s post hoc test). F, G Electrophysiology analysis. ApoE4 overexpression worsened the LTP defects in wild-type or 3xTg mice. LTP of fEPSPs (mean ± SEM; n = 6 in each group; *P < 0.05, **P < 0.01 compared with group1, two-way ANOVA and Bonferroni’s post hoc test). Shown traces were representative fEPSPs of ten samples recorded before and after TBS (theta-burst stimulation). H Quantification of fEPSP potentiation from the final 10 min of recordings (86–95 min in Fig. 4G, H) normalized to basal levels. Representative recording traces: black line, baseline; red line, LTP 86–95 min (mean ± SEM; n = 6 in each group; *P < 0.05 compared with 3xTg-control, two-way ANOVA and Bonferroni’s post hoc test). I, J Morris water maze analysis. ApoE4 overexpression exacerbated the learning and memory dysfunctions (mean ± SEM; n = 7–8 mice per group; *P < 0.05 compared with group1, two-way ANOVA and Bonferroni’s post hoc test). K, L Fear condition tests. Contextual and cued fear conditions were reduced in ApoE overexpressed mice. (Mean ± SEM; n = 7–8 mice per group; *P < 0.05, **P < 0.01 compared with group1, two-way ANOVA and Bonferroni’s post hoc test). See also Supplementary Fig. 11.

Article Snippet: Differentiation of human iPSC-derived NSCs into neurons The used human induced pluripotent stem cell (iPSC)-derived NSCs were obtained from two donors: ax0111 from AD patient with ApoE4/4 genotype, ax0112 from AD patient with ApoE3/3 genotype (Axol Bioscience, Cambridge, UK).

Techniques: Injection, Western Blot, Over Expression, Activity Assay, Expressing, Staining

Journal: Molecular Psychiatry

Article Title: C/EBPβ is a key transcription factor for APOE and preferentially mediates ApoE4 expression in Alzheimer’s disease

doi: 10.1038/s41380-020-00956-4

Figure Lengend Snippet: A AD patients with ApoE4/4 genotype showed higher CEBPB and APOE mRNA level than ApoE3/3. Data represented 4–5 independent experiments (mean ± SEM, **P < 0.01, two-tailed student’s t test). B The AD patients’ brain cortex tissues were analyzed by immunoblotting. C/EBPβ and its active p-C/EBPβ T235 signals were selectively escalated in ApoE4/4 AD brains versus ApoE3/3 brains. C Quantitation of western blot data. Data represent mean ± SEM (n = 4–5, *P < 0.05, **P < 0.01, two-tailed student’s t test). D The human iPSC-derived neurons from ApoE3/3 or ApoE4/4 genotype were stained for NeuN (red) and MAP2 (green) after cultured for 28 days, scale bar: 40 μm. E, F The human iPSC-derived neurons were treated with lentivirus overexpressing or knocking down C/EBPβ for 7 days, respectively. CEBPB and APOE mRNA levels in induced human neurons were detected by qPCR (mean ± SEM, n = 3, *P < 0.05, **P < 0.01, two-way ANOVA and Bonferroni’s post hoc test) (E), and ApoE and AEP protein levels were detected by western blot (F). G Quantitation of western blot data. Data represent mean ± SEM (n = 3, *P < 0.05, **P < 0.01, two-way ANOVA and Bonferroni’s post hoc test).

Article Snippet: Differentiation of human iPSC-derived NSCs into neurons The used human induced pluripotent stem cell (iPSC)-derived NSCs were obtained from two donors: ax0111 from AD patient with ApoE4/4 genotype, ax0112 from AD patient with ApoE3/3 genotype (Axol Bioscience, Cambridge, UK).

Techniques: Two Tailed Test, Western Blot, Quantitation Assay, Derivative Assay, Staining, Cell Culture

Journal: iScience

Article Title: Imaging lipid rafts reveals the principle of ApoE4-induced Aβ upregulation in human neurons

doi: 10.1016/j.isci.2025.111893

Figure Lengend Snippet:

Article Snippet: Human iPSC line from a healthy control ( APOE3 ) , The Jackson Laboratory , Cat#JIPC1000.

Techniques: Recombinant, Membrane, Electron Microscopy, Enzyme-linked Immunosorbent Assay, Conjugation Assay, Control, Software

Journal: Stem Cell Reports

Article Title: APOE4 -carrying human astrocytes oversupply cholesterol to promote neuronal lipid raft expansion and Aβ generation

doi: 10.1016/j.stemcr.2021.07.017

Figure Lengend Snippet: Extracellular cholesterol positively regulates the formation of lipid rafts and its association with APP in hiPSC-derived neurons (A) Measurement of secretory levels of cholesterols in ApoE3 or ApoE4 ACM. (B) Filipin III signals in ACM (n = 4 experiments). (C) Levels of total cholesterol, free cholesterol, and cholesteryl ester in ACM (n = 3 experiments). (D) Images of filipin III staining in neurons. Scale bar, 10 μm. Right: quantification of filipin III area, intensity, and area × intensity in neurons (n = 13–15 images from three experiments). (E) Images of CTX-B and APP staining in neurons. Scale bar, 10 μm. (F) CTX-B area, intensity, and area × intensity in neurons (n = 12 images from three experiments). (G) APP area, intensity, and area × intensity in neurons (n = 12 images from three experiments). (H) Co-localization of CTX-B and APP in neurons (n = 12 images from three experiments). (I) APP intensity in CTX-B/APP co-localization area in neurons (n = 12 images from three experiments). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant (Student's t test or ANOVA followed by Dunnett's post hoc test). Error bars represent SEM.

Article Snippet: The ApoE3 iPSC line was generated from the Coriell Institute's fibroblast line derived from a healthy individual (age 75 years, female; #AG09173) by Dr. Yankner’s Laboratory at Harvard Medical School ( ).

Techniques: Derivative Assay, Staining