endogenous mouse apoe gene  (Inotiv)

Journal: iScience

Article Title: APOE4 drives widespread changes to the hepatic proteome and alters metabolic function

doi: 10.1016/j.isci.2026.115035

Figure Lengend Snippet: Whole liver proteomics in female and male APOE3 and APOE4 mice (A) Study schematic showing primary outcomes in APOE -targeted replacement (TR) mice. n = 3 mice per group for all mouse proteomic outcomes. (B) APOE protein expression from whole liver measured via ELISA in male and female APOE3 and APOE4 mice. Statistical significance was determined by two-way ANOVA. Data are presented as mean ± SD. n = 8 mice per genotype and sex. (C) The number of differentially expressed (DE) proteins in male and female APOE3 and APOE4 mice. (D) Volcano plot showing upregulated and downregulated proteins in female APOE4 vs. APOE3 mice. (E) IPA pathway showing top 5 upregulated and downregulated pathways in female APOE4 vs. APOE3 mice. (F) Volcano plot showing upregulated and downregulated proteins in male APOE4 vs. APOE3 mice. (G) IPA pathway analysis showing top 5 upregulated and downregulated pathways in male APOE4 vs. APOE3 mice. (H) Heatmap of proteins involved in fatty acid metabolism, cholesterol and bile acid metabolism, lipid transport, and lipid storage showing upregulated and downregulated proteins between APOE4 and APOE3 mice. (I) Venn diagram showing the number of shared upregulated proteins between comparisons of female APOE4 vs. APOE3 mice and male APOE4 vs. APOE3 mice. (J) Venn diagram showing the number of downregulated proteins shared between comparisons of female APOE4 vs. APOE3 mice and male APOE4 vs. APOE3 mice. (K) Violin plot showing normalized Pgam1 protein expression from liver proteomics in male and female APOE3 and APOE4 mice. Statistical significance was determined by two-way ANOVA with Fisher’s LSD post hoc. ∗ p < 0.05. G, genotype. n = 3 mice per genotype and sex. See also .

Article Snippet: These APOE TR mice were previously generated by removing and replacing the endogenous mouse Apoe gene with the human APOE gene., Mice were maintained on a standard chow diet (Teklad Global Rodent Diet, 8604) for the entire study and housed at ∼25°C with a standard 12-h light/dark cycle.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: APOE4 drives widespread changes to the hepatic proteome and alters metabolic function

doi: 10.1016/j.isci.2026.115035

Figure Lengend Snippet: Proteomics analysis of APOE isogenic iHLCs (A) iHLC study schematic. Two pairs of isogenic iPSCs (A and B) homozygous for either APOE3 or APOE4 were used to generate iHLCs and examine proteomic and bioenergetic outcomes. n = 5 per group for all iHLC proteomic outcomes. (B) Violin plots of APOE protein expression between two sample batches in pair A isogenics. Statistical significance was determined by unpaired t test. ns, not significant, ∗∗∗ p < 0.001. (C) Volcano plot showing upregulated and downregulated proteins between APOE4 and APOE3 iHLCs from batch 1. (D) Volcano plot showing upregulated and downregulated proteins between APOE4 and APOE3 iHLCs from batch 2. (E) Heatmap of the top 20 most significant upregulated and top 20 most significant downregulated proteins ( p < 0.05) shared between batch 1 and batch 2. (F) IPA pathway analysis showing top 12 upregulated and top 12 downregulated pathways shared in both batch 1 and 2 between APOE4 and APOE3 iHLCs. (G) STRING network analysis of the most significant upregulated and downregulated proteins shared between batch 1 and batch 2 in APOE4 vs. APOE3 iHLCs. (H) Cellular component Gene Ontology (GO) analysis of the top 20 upregulated and top 20 downregulated proteins in APOE4 vs. APOE3 iHLCs. See also .

Article Snippet: These APOE TR mice were previously generated by removing and replacing the endogenous mouse Apoe gene with the human APOE gene., Mice were maintained on a standard chow diet (Teklad Global Rodent Diet, 8604) for the entire study and housed at ∼25°C with a standard 12-h light/dark cycle.

Techniques: Expressing

Journal: iScience

Article Title: APOE4 drives widespread changes to the hepatic proteome and alters metabolic function

doi: 10.1016/j.isci.2026.115035

Figure Lengend Snippet: Mitochondrial function in APOE isogenic iHLCs (A) Mitochondrial stress test (MST) tracing from isogenic pair A. (B) Quantification of basal respiration, maximal respiration, proton leak, and ATP-production linked respiration from isogenic pair an MST. n = 13–14 per group. (C) MST tracing from isogenic pair B. (D) Quantification of basal respiration, maximal respiration, proton leak, and ATP-production-linked respiration from isogenic pair B MST. n = 12–15 per group. (E) Electron transport chain (ETC) flux through complex I, II, III, and IV in isogenic pair A. n = 7–22 per group. (F) ETC flux through complex I, II, III, and IV in isogenic pair B. n = 14–22 per group. (G) Representative tetramethyl rhodamine, ethyl ester, perchlorate (TMRE) images from isogenic pair A. Scale bars, 50 μm. (H–K) Quantification of TMRE (H), MitoSOX (I), Amplex Red (J), and Rhod-2 AM (K) fluorescence intensities from isogenic pair A. n = 8–16 per group. (L) Representative TMRE images from isogenic pair B. Scale bars, 50 μm. (M–P) Quantification of TMRE (M), MitoSOX (N), Amplex Red (O), and Rhod-2 AM (P) fluorescence intensities from isogenic pair B. n = 8–16 per group. (B, D, E–F, H–K, and M–P) Statistical significance was determined by unpaired t test. Data are shown as mean ± SD. ns = not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: These APOE TR mice were previously generated by removing and replacing the endogenous mouse Apoe gene with the human APOE gene., Mice were maintained on a standard chow diet (Teklad Global Rodent Diet, 8604) for the entire study and housed at ∼25°C with a standard 12-h light/dark cycle.

Techniques: Fluorescence

Journal: iScience

Article Title: APOE4 drives widespread changes to the hepatic proteome and alters metabolic function

doi: 10.1016/j.isci.2026.115035

Figure Lengend Snippet: Glycolytic function and glucose/pyruvate oxidation in APOE isogenic iHLCs (A) Glycolytic stress test (GST) tracing from isogenic pair A. (B) Quantification of basal glycolysis, glycolytic capacity, glycolytic reserve, and non-glycolytic acidification from isogenic pair A GST. n = 11 per group. (C) GST tracing from isogenic pair B. (D) Quantification of basal glycolysis, glycolytic capacity, glycolytic reserve, and non-glycolytic acidification from isogenic pair B GST. n = 10–16 per group. (E) Glucose/pyruvate oxidation stress test from isogenic pair A. (F) Quantification of basal respiration, acute response, and maximal respiration from isogenic pair A glucose/pyruvate oxidation stress test. n = 7–10 per group. (G) Glucose/pyruvate oxidation stress test from isogenic pair B. (H) Quantification of basal respiration, acute response, and maximal respiration from isogenic pair B glucose/pyruvate oxidation stress test. n = 7–11 per group. (I) Significant DE proteins involved in glucose metabolism pathway from IPA. (J) Glucose metabolism protein network from STRING analysis. (B and D) Statistical significance was determined by unpaired t test and (F and H) two-way ANOVA with Fisher’s LSD post hoc. Data are shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. D, drug, G, genotype.

Article Snippet: These APOE TR mice were previously generated by removing and replacing the endogenous mouse Apoe gene with the human APOE gene., Mice were maintained on a standard chow diet (Teklad Global Rodent Diet, 8604) for the entire study and housed at ∼25°C with a standard 12-h light/dark cycle.

Techniques:

Journal: iScience

Article Title: APOE4 drives widespread changes to the hepatic proteome and alters metabolic function

doi: 10.1016/j.isci.2026.115035

Figure Lengend Snippet: Fatty acid oxidation and LipidTOX staining in APOE isogenic iHLCs (A) Long chain fatty acid (LCFA) oxidation stress test from isogenic pair A. (B) Quantification of basal respiration, acute response, and maximal respiration from isogenic pair An LCFA oxidation stress test. n = 6–8 per group. (C) LCFA oxidation stress test from isogenic pair B. (D) Quantification of basal respiration, acute response, and maximal respiration from isogenic pair B LCFA oxidation stress test. n = 7–10 per group. (E) Representative LipidTOX images from isogenic pair A. Scale bars, 50 μm. (F) Quantification of lipid droplet (LD) numbers per cell and diameter (μm) from isogenic pair A. n = 24 per group. (G) Representative LipidTOX images from isogenic pair B. Scale bars, 50 μm. (H) Quantification of LD numbers per cell and diameter (μm) from isogenic pair B. n = 24 per group. (I) Comparison analysis showing significantly altered proteins in SREBF2 network from proteomics. (B and D) Statistical significance was determined by a two-way ANOVA with Fisher’s LSD post hoc and (F and H) unpaired t test. Data are shown as mean ± SD. ns = not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. D, drug, G, genotype.

Article Snippet: These APOE TR mice were previously generated by removing and replacing the endogenous mouse Apoe gene with the human APOE gene., Mice were maintained on a standard chow diet (Teklad Global Rodent Diet, 8604) for the entire study and housed at ∼25°C with a standard 12-h light/dark cycle.

Techniques: Staining, Comparison

Journal: Molecular neurodegeneration

Article Title: Sex-dependent calcium hyperactivity due to lysosomal-related dysfunction in astrocytes from APOE4 versus APOE3 gene targeted replacement mice.

doi: 10.1186/s13024-020-00382-8

Figure Lengend Snippet: Fig. 5 Effects of the APOE allele vs APOE contents on Ca2+-based excitability. a APOE mRNA expression relative to Gapdh and 18S mRNA contents in APOE3 and APOE4 immortalized astrocytes, quantified by real time qPCR (N = 3). b Representative ApoE western blot of APOE3 cells transfected with scramble or APOE siRNA. c Representative traces and quantification of the magnitude of ATP-induced Ca2+ responses in APOE3 astrocytes transfected with lipofectamine (lipo), plus scramble (sc) or APOE siRNA, and APOE4 astrocytes treated only with lipofectamine (N = 4). d Representative images of immunocytochemistry of APOE3 and APOE4 cells transfected with plasmids expressing GFP, GFP-APOE4, and GFP-APOE3, as indicated. Scale bar represents 15 μm. e Representative traces and quantification of the magnitude of 100 μM ATP-elicited Ca2+ responses in APOE3 and APOE4 cells transfected with different plasmids as indicated. Ca2+ peaks after ATP stimulation are relative to the responses of APOE3 cells transfected with GFP. Only GFP fluorescent cells were analyzed (at least 30 cells from 4 independent experiments). f Quantification of the magnitude of Ca2+ responses elicited by 100 μM ATP in APOE3 astrocytes transfected with GFP or GFP-APOE4 plasmids, as indicated. Ca2+ peaks after ATP stimulation are relative to the responses of APOE3 cells transfected with GFP. Only GFP fluorescent cells were analyzed (at least 30 cells from 4 independent experiments). g Representative images of Lamp1 (red) and ApoE (green) immunocytochemistry in APOE3 cells. Merged images and amplifications of the area in the white frame, are displayed. Scale bar represents 15 μm. h Representative images of Lamp1 immunocytochemistry (red) and GFP-ApoE fluorescence (green) in APOE4 cells. Merged images and the amplification of a cell of each image, indicated with a white frame, are displayed. Scale bar represents 15 μm. Unpaired parametric T-test was used in a, and one-way ANOVA in c and e. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****)

Article Snippet: Nine- to 12-week-old male and female APOE3 and APOE4 transgenic mice homozygous for the human APOE3 or APOE4 gene replacing the endogenous mouse APOE gene were purchased from Taconic (USA) [42, 43].

Techniques: Expressing, Western Blot, Transfection, Immunocytochemistry, Fluorescence, Amplification

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Human Apolipoprotein E Isoforms differentially affect Bone Mass and Turnover in vivo

doi: 10.1002/jbmr.1757

Figure Lengend Snippet: APOE genotype distribution, age, BMI and smoking status in the NPHS II subgroup analyzed in this study.

Article Snippet: Transgenic mice, homozygous for targeted replacement of the mouse endogenous Apoe gene with the human APOE2 , APOE3 and APOE4 on a C57BL/6 background were purchased from Taconic ( www.taconic.com ).

Techniques:

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Human Apolipoprotein E Isoforms differentially affect Bone Mass and Turnover in vivo

doi: 10.1002/jbmr.1757

Figure Lengend Snippet: The concentrations of the bone turnover markers bone-specific alkaline phosphatase (BSAP), osteocalcin (OCN), N-terminal telopeptide of collagen type 1 (NTX) (A) as well as osteoprotegerin (OPG) and RANK ligand (RANKL) (B) were determined with standard procedures from serum specimens of fasted male individuals from the NPHSII study (compare Table 1). The bars represent the geometric mean values +/− SD. (APOE ε2ε2 n=21; APOE ε3ε3 n=80; APOE ε4ε4 n=55). Statistical differences were calculated by paired or unpaired t-test *p<0.05, **p<0.01, ***p<0.001

Article Snippet: Transgenic mice, homozygous for targeted replacement of the mouse endogenous Apoe gene with the human APOE2 , APOE3 and APOE4 on a C57BL/6 background were purchased from Taconic ( www.taconic.com ).

Techniques: