Journal: International Journal of Molecular Sciences

Article Title: Sex and APOE Genotype Alter the Basal and Induced Inflammatory States of Primary Microglia from APOE Targeted Replacement Mice

doi: 10.3390/ijms23179829

Figure Lengend Snippet: Basal pro− and anti−inflammatory gene expression in mixed−sex microglia is APOE genotype−dependent: ( A ) basal pro−inflammatory and ( B ) anti−inflammatory gene expression was evaluated in non−stimulated mixed−sex microglia from APOE3 and APOE4 genotypes. Gene expression between genotypes was analyzed using an unpaired Student’s t −test (N = 3 isolations, n = 6 plates). Data are represented as the mean ± standard deviation. *, ***, and **** indicate statistical significance for the comparison and denote p < 0.05, 0.001, and 0.0001, respectively. ns = not statistically significant ( p > 0.05).

Article Snippet: TR-APOE breeder mice with a C57BL/6 background, with the mouse APOE gene replaced by a homozygous human APOE3 or APOE4 gene [ ], were purchased from Taconic and bred in-house.

Techniques: Gene Expression, Standard Deviation, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Sex and APOE Genotype Alter the Basal and Induced Inflammatory States of Primary Microglia from APOE Targeted Replacement Mice

doi: 10.3390/ijms23179829

Figure Lengend Snippet: Comparison of gene expression between APOE3 and APOE4 genotypes: ( A ) pro− and ( B ) anti−inflammatory gene expression was evaluated in mixed−sex microglia from APOE3 and APOE4 genotypes following treatment with LPS (10 ng/mL) and LPS (10 ng/mL) + IFNg (10 ng/mL) for 6 h. Gene expression between genotypes was assessed by two−way ANOVA followed by Tukey’s multiple comparisons test (N = 3 isolations, n = 6 plates). Each data point represents the average mRNA expression from one isolation. Data are represented as the mean ± standard deviation. # denotes a significant increase compared to the respective genotype control (data not included in the graphs, p < 0.05). *, **, ***, and **** indicate statistical significance for the comparison and denote p < 0.05, 0.01, 0.001, and 0.0001, respectively. ( C ) Heat map comparing the mean expression of pro− and anti−inflammatory genes in control, LPS, and LPS + IFNg treated APOE3 and APOE4 PMG. * and ^ denote a significant ( p < 0.05) increase and decrease, respectively, in APOE4 gene expression compared to APOE3.

Article Snippet: TR-APOE breeder mice with a C57BL/6 background, with the mouse APOE gene replaced by a homozygous human APOE3 or APOE4 gene [ ], were purchased from Taconic and bred in-house.

Techniques: Comparison, Gene Expression, Expressing, Isolation, Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Sex and APOE Genotype Alter the Basal and Induced Inflammatory States of Primary Microglia from APOE Targeted Replacement Mice

doi: 10.3390/ijms23179829

Figure Lengend Snippet: APOE4 genotype exacerbates release of pro− and anti−inflammatory cytokines: cytokine levels were measured following treatment with LPS or LPS + IFNg for 24 h. ( A ) Average levels of nitrite (±standard deviation) accumulated in the media were measured by GRIESS assay. Using two−way ANOVA, significant treatment and genotype effects were observed. # denotes a significant increase compared to the respective genotype control ( p < 0.05). ( B ) Basal levels of secreted cytokines were measured in non−stimulated microglia after 24 h by an MSD multiplex assay. Comparisons between genotypes were made using an unpaired Student’s t −test. ( C ) Cytokine levels in the media were measured after treatment with LPS or a combination of LPS and IFNg for 24 h. Two−way ANOVA was used to assess cytokine release in mixed−sex microglia from APOE3 and APOE4 genotypes (N = 3−4 isolations, n = 6−8 plates). Data are represented as the mean ±standard deviation, and each data point denotes the average of each experimental plate. # denotes a significant increase compared to the respective genotype control (data not included in the graphs, p < 0.05). *, **, ***, and **** indicate statistical significance for the comparison and denote p < 0.05, 0.01, 0.001, and 0.0001, respectively. ns = not statistically significant ( p >0.05).

Article Snippet: TR-APOE breeder mice with a C57BL/6 background, with the mouse APOE gene replaced by a homozygous human APOE3 or APOE4 gene [ ], were purchased from Taconic and bred in-house.

Techniques: Standard Deviation, Griess Assay, Control, Multiplex Assay, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Sex and APOE Genotype Alter the Basal and Induced Inflammatory States of Primary Microglia from APOE Targeted Replacement Mice

doi: 10.3390/ijms23179829

Figure Lengend Snippet: Basal pro− and anti−inflammatory gene expression in sex−specific microglia is dependent on both the APOE genotype and sex: ( A ) basal pro−inflammatory and ( B ) anti−inflammatory gene expression was evaluated in non−stimulated sex−specific microglia from APOE3 and APOE4 genotypes. Gene expression between genotypes and sexes was analyzed using two−way ANOVA (N = 3 isolations, n = 6 plates). Data are represented as the mean ± standard deviation. *, **, ***, and **** indicate statistical significance for the comparison and denote p < 0.05, 0.01, 0.001, and 0.0001, respectively. ns = not statistically significant ( p > 0.05).

Article Snippet: TR-APOE breeder mice with a C57BL/6 background, with the mouse APOE gene replaced by a homozygous human APOE3 or APOE4 gene [ ], were purchased from Taconic and bred in-house.

Techniques: Gene Expression, Standard Deviation, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Sex and APOE Genotype Alter the Basal and Induced Inflammatory States of Primary Microglia from APOE Targeted Replacement Mice

doi: 10.3390/ijms23179829

Figure Lengend Snippet: Gene expression in sex-specific microglia is dependent on both the APOE genotype and sex: ( A ) pro− and ( B ) anti−inflammatory gene expression was evaluated in sex−specific microglia from APOE3 and APOE4 genotypes following LPS or LPS + IFNg treatment for 6 h. Gene expression between genotypes and sexes was analyzed using a three−way ANOVA (N = 3 isolations, n = 6 plates) followed by Tukey’s multiple comparisons test. Each data point represents the average mRNA expression from one isolation. Data are represented as the mean ± standard deviation. # denotes a significant increase compared to the respective genotype control (data not included in the graphs, p < 0.05). *, **, ***, and **** indicate statistical significance for the comparison and denote p < 0.05, 0.01, 0.001, and 0.0001, respectively. ns = not statistically significant ( p >0.05). ( C ) Heat map comparing the mean expression of pro− and anti−inflammatory genes in sex−specific APOE3 and APOE4 PMG treated with LPS and LPS + IFNg. * and ^ denote a significant ( p < 0.05) increase and decrease, respectively, in fold changes of gene expression compared to a genotype−matched male PMG.

Article Snippet: TR-APOE breeder mice with a C57BL/6 background, with the mouse APOE gene replaced by a homozygous human APOE3 or APOE4 gene [ ], were purchased from Taconic and bred in-house.

Techniques: Gene Expression, Expressing, Isolation, Standard Deviation, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Sex and APOE Genotype Alter the Basal and Induced Inflammatory States of Primary Microglia from APOE Targeted Replacement Mice

doi: 10.3390/ijms23179829

Figure Lengend Snippet: APOE genotype and sex-dependent inflammation are mediated through p65 activation: ( A ) representative images for immunocytochemistry of p65 in control and LPS + IFNg treated APOE3 and APOE4, male and female PMG (scale bar = 100 μm). ( B ) Quantification of total p65 and ( C ) quantification of nuclear p65. N = 3 isolations. Each data point denotes the average p65 integrated density/cell density for an individual isolation. A denotes a significant increase compared to the E3 male control ( p < 0.05). # denotes a significant increase compared to the respective genotype and sex control ( p < 0.05). Data are represented as the mean ± standard deviation. * and **** indicate statistical significance for the comparison and denote p < 0.05 and 0.0001, respectively. ns = not statistically significant ( p > 0.05).

Article Snippet: TR-APOE breeder mice with a C57BL/6 background, with the mouse APOE gene replaced by a homozygous human APOE3 or APOE4 gene [ ], were purchased from Taconic and bred in-house.

Techniques: Activation Assay, Immunocytochemistry, Control, Isolation, Standard Deviation, Comparison

Journal: The FASEB Journal

Article Title: Altered SPMs and age‐associated decrease in brain DHA inAPOE4female mice

doi: 10.1096/fj.201900423r

Figure Lengend Snippet: Figure 1. DHA (22:6 n-3) and oleic acid (18:1 n-9) as a percentage of total fatty acids in the cortex (A, B) and hippocampus (C, D) of female and male APOE3 and APOE4 mice at 2, 9, and 18 mo of age fed on a chow diet. Values are means 6 SEM; n = 6–7/group. **P , 0.01, ***P , 0.001.

Article Snippet: In total, 120 male and female humanized APOE3 [B6.129P2Apoetm2(APOE*3)Mae N8]- and APOE4 [B6.129P2-Apoetm2(APOE*4)Mae N8]-targeted replacement mice homozygous for the human APOE3 orAPOE4 gene (Taconic Farms, Germantown,NY,USA) were used in these experiments.

Techniques:

Journal: The FASEB Journal

Article Title: Altered SPMs and age‐associated decrease in brain DHA inAPOE4female mice

doi: 10.1096/fj.201900423r

Figure Lengend Snippet: Figure 2. DHA-containing PL fractions in cortex of female and male APOE3 and APOE4 mice at 2 and 18 mo of age fed a chow diet. The age*genotype interaction was only significant for PE-DHA (P = 0.005; D). G–I) Figures show DHA-containing PE, PS, and PC, respectively, at a group level in which PE-DHA content was significantly decreased in female APOE4 at 18 mo (P , 0.001). Values are means 6 SEM; n = 15–16/group for age effect (A–C), n = 7–8/ group for age*genotype interaction (D–F), n = 3–4/group for group effect (G–I). Inclusion of arrows indicates significant interaction. ***P , 0.001.

Article Snippet: In total, 120 male and female humanized APOE3 [B6.129P2Apoetm2(APOE*3)Mae N8]- and APOE4 [B6.129P2-Apoetm2(APOE*4)Mae N8]-targeted replacement mice homozygous for the human APOE3 orAPOE4 gene (Taconic Farms, Germantown,NY,USA) were used in these experiments.

Techniques:

Journal: The FASEB Journal

Article Title: Altered SPMs and age‐associated decrease in brain DHA inAPOE4female mice

doi: 10.1096/fj.201900423r

Figure Lengend Snippet: Figure 3. Ratios of DHA:AA in total lipid and individual PL species in the cortex of female and male APOE3 and APOE4 mice at 2 and 18 mo of age fed a chow diet. The age*genotype interaction was significant for total DHA:AA ratio (P = 0.01; B), PE-DHA:AA (P = 0.00; E), and PC-DHA:AA (P = 0.04; K). Values are means 6 SEM; n = 15–27/group for age effect (A, D, G, J), n = 7–14/ group for age*genotype interaction (B, E, H, K), n = 4–7/group for group effect (C, F, I, L). Inclusion of arrows indicates significant interaction. *P = 0.01, **P , 0.01, ***P , 0.001.

Article Snippet: In total, 120 male and female humanized APOE3 [B6.129P2Apoetm2(APOE*3)Mae N8]- and APOE4 [B6.129P2-Apoetm2(APOE*4)Mae N8]-targeted replacement mice homozygous for the human APOE3 orAPOE4 gene (Taconic Farms, Germantown,NY,USA) were used in these experiments.

Techniques:

Journal: The FASEB Journal

Article Title: Altered SPMs and age‐associated decrease in brain DHA inAPOE4female mice

doi: 10.1096/fj.201900423r

Figure Lengend Snippet: Figure 4. Sex differences in the ratios of SPMs to their parent compound, EPA or DHA, in cortex of female and male APOE3 and APOE4 mice at 2, 9, and 18 mo of age fed a chow diet. Figures of SPMs and their precursors were expressed in pg/mg of tissue to calculate the ratios. Values are means 6 SEM; n = 20/group. *P # 0.05, **P , 0.01, ***P , 0.001.

Article Snippet: In total, 120 male and female humanized APOE3 [B6.129P2Apoetm2(APOE*3)Mae N8]- and APOE4 [B6.129P2-Apoetm2(APOE*4)Mae N8]-targeted replacement mice homozygous for the human APOE3 orAPOE4 gene (Taconic Farms, Germantown,NY,USA) were used in these experiments.

Techniques:

Journal: The FASEB Journal

Article Title: Altered SPMs and age‐associated decrease in brain DHA inAPOE4female mice

doi: 10.1096/fj.201900423r

Figure Lengend Snippet: Figure 6. Expression level of 2 proinflammatory cytokines, IL-6 and TNF-a, in the cortex of female and male APOE3 and APOE4 mice at 2, 9, and 18 mo of age fed a chow diet. Values are means 6 SEM; n = 11–12/group (A, D) and n = 17–18/group for sex (B, E) or genotype effect (C, F).

Article Snippet: In total, 120 male and female humanized APOE3 [B6.129P2Apoetm2(APOE*3)Mae N8]- and APOE4 [B6.129P2-Apoetm2(APOE*4)Mae N8]-targeted replacement mice homozygous for the human APOE3 orAPOE4 gene (Taconic Farms, Germantown,NY,USA) were used in these experiments.

Techniques: Expressing

Journal: The FASEB Journal

Article Title: Altered SPMs and age‐associated decrease in brain DHA inAPOE4female mice

doi: 10.1096/fj.201900423r

Figure Lengend Snippet: Figure 5. Genotype differences in the ratios of SPMs to their parent compound, EPA or DHA, in cortex of female and male APOE3 and APOE4 mice at 2, 9, and 18 mo of age fed a chow diet. Figures of SPMs and their precursors were expressed in pg/mg of tissue to calculate the ratios. Values are means 6

Article Snippet: In total, 120 male and female humanized APOE3 [B6.129P2Apoetm2(APOE*3)Mae N8]- and APOE4 [B6.129P2-Apoetm2(APOE*4)Mae N8]-targeted replacement mice homozygous for the human APOE3 orAPOE4 gene (Taconic Farms, Germantown,NY,USA) were used in these experiments.

Techniques:

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: a ApoJ protein levels in serum of liver-specific ApoJ-deficient mice (L-ApoJ −/− ). Serum was separated by SDS–PAGE. ApoJ or ApoB 48 was visualized by immunoblotting. These data are representative from more than three independent experiments. b ApoJ protein levels in multiple metabolic organs of L-ApoJ −/− . Tissue lysates (20–50 μg) were separated by SDS–PAGE. ApoJ or actin was visualized by immunoblotting. These data are representative from more than three independent experiments. Mus muscle, Adi adipose tissue, Hypo hypothalamus, Panc pancreas, Kid kidney, Stom stomach, Intes intestine, Spl spleen, BAT brown adipose tissue; C: ApoJ loxP/loxP mice, KO: albumin-Cre: ApoJ loxP/loxP mice (L-ApoJ −/− ). c mRNA levels of ApoJ in multiple metabolic organs of L-ApoJ −/− mice. mRNA levels of ApoJ were measured by RT-PCR. Mice were studied at 12 weeks of age. Molecular marker (MM) is 100 bp DNA ladder. 313 bp indicates ApoJ mRNA. These data are representative from three independent experiments; Con: ApoJ loxP/loxP mice, KO: albumin-Cre: ApoJ loxP/loxP mice (L-ApoJ −/− ).

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: SDS Page, Western Blot, Reverse Transcription Polymerase Chain Reaction, Marker

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: a ApoJ protein levels in serum of L-ApoJ −/− mice injected with ApoJ or GFP adenovirus. L-ApoJ −/− or global ApoJ −/− mice were injected with a recombinant adenovirus encoding a secretory ApoJ or GFP at a concentration of 2 × 0 9 pfu per gram of body weight via the tail vein. A: ApoJ-expressing adenovirus, G: GFP-expressing adenovirus. Serum was separated by SDS–PAGE. ApoJ, ApoB 48 or actin were visualized by immunoblotting. b ApoJ protein levels in multiple metabolic organs of in L-ApoJ −/− mice injected with ApoJ or GFP adenovirus. Tissue lysates (20–50 μg) were separated by SDS–PAGE. ApoJ or GAPDH were visualized by immunoblotting. c ApoJ protein levels in serum of in global ApoJ −/− mice injected with ApoJ or GFP adenovirus. d ApoJ protein levels in multiple metabolic organs of global ApoJ −/− mice. Tissue lysates (20–50 μg) were separated by SDS–PAGE. All data are representative from three independent experiments. Mice were studied at 9–10 weeks of age. Hypo hypothalamus, BAT brown adipose tissue.

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: Injection, Recombinant, Concentration Assay, Expressing, SDS Page, Western Blot

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: a ApoJ protein levels in serum of muscle-specific ApoJ-deficient mice (M-ApoJ −/− ). Serum was separated by SDS–PAGE. ApoJ was visualized by immunoblotting. b ApoJ protein levels in multiple metabolic organs of M-ApoJ −/− . Tissue lysates (20–50 μg) were separated by SDS–PAGE. ApoJ was visualized by immunoblotting. c mRNA levels of ApoJ in multiple metabolic organs of M-ApoJ −/− mice. mRNA levels of ApoJ were measured by RT-PCR. Mice were studied at 8 weeks of age. Molecular marker (MM) is 100 bp DNA ladder. 313 bp indicates ApoJ mRNA. All data are representative from three independent experiments. Con: ApoJ loxP/loxP mice, KO: myogenin-Cre: ApoJ loxP/loxP mice (M-ApoJ −/− ).

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: SDS Page, Western Blot, Reverse Transcription Polymerase Chain Reaction, Marker

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: a Body weight ( n = 7 for control, n = 9 for L-ApoJ −/− ), b fasting blood glucose ( n = 7 for control, n = 9 for L-ApoJ −/− ), c fasting serum insulin ( n = 8 per group), d serum IGF-1 ( n = 10 for control, n = 9 for L-ApoJ −/− ), e serum IGFBP-1 ( n = 10 for control, n = 9 for L-ApoJ −/− ), f plasma IGFBP-3 ( n = 11 for control, n = 12 for L-ApoJ −/− ), g serum triglycerides ( n = 6 for control, n = 7 for L-ApoJ −/− ), h serum cholesterol ( n = 6 for control, n = 7 for L-ApoJ −/− ), i liver triglycerides ( n = 7 per group), j liver cholesterol ( n = 6 per group), k H&E-stained liver sections, l hepatic VDLD-TG production ( n = 5 per group), m results of insulin tolerance test (ITT; n = 6 per group), n results of glucose tolerance test (GTT; n = 6 for control, n = 8 for L-ApoJ −/− ), o glucose-stimulated insulin secretion test (GSIS; n = 3 for control, n = 4 for L-ApoJ −/− ), p muscle insulin signaling, q liver insulin signaling, and r adipose tissue insulin signaling ( n = 7 per group) were measured in control and L-ApoJ −/− mice. Serum parameters were measured from overnight fasted mice at 11–13 weeks of age. H&E-stained liver sections are representative from three independent experiments. The scale bars represent 100 μm. ITT was performed at 21 weeks of age and GTT was performed at 13 weeks of age. GSIS was performed at 16–18 weeks of age. Insulin signaling was performed at 25–26 weeks of age; AUC area under the curve. Logarithmic transformation was done for GTT. The AUC in GSIS was calculated by 30 min. All bars and errors represent means ± SEM. p values by two-sided Student’s t -test in b and e , and by repeated measures two-way ANOVA in m and n are indicated. p values for AUCs were evaluated by one-sided Student’s t -test for m and n , and by Mann–Whitney U test for o based on the distribution of the data. * p < 0.05 vs control by repeated measures two-way ANOVA.

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: Control, Clinical Proteomics, Staining, Transformation Assay, MANN-WHITNEY

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: a ApoJ protein levels in serum of muscle-specific LRP2-deficient mice (M-LRP2 −/− ). Serum ApoJ levels were measured by ELISA. ApoJ, or ApoB 48 were visualized by immunoblotting. Scatter diagram shows densitometric quantitation of ApoJ protein from control and M-LRP2 −/− mice ( n = 7 per group). b ApoJ protein levels in muscle of M-LRP2 −/− . Muscle lysates (20 μg) were separated by SDS–PAGE. ApoJ or GAPDH was visualized by immunoblotting. Scatter diagram shows densitometric quantitation of ApoJ protein from control and M-LRP2 −/− mice ( n = 8 per group). c ApoJ protein levels in multiple metabolic organs of M-LRP2 −/− . Tissue lysates (20–50 μg) were separated by SDS–PAGE. ApoJ was visualized by immunoblotting. d Body weight ( n = 12 per group), e random blood glucose ( n = 10 per group), f serum insulin ( n = 11 for control, n = 7 for M-LRP2 −/− ), g insulin tolerance test (ITT; n = 5 per group), h glucose tolerance test (GTT; n = 5 per group), i glucose-stimulated insulin secretion test (GSIS; n = 4 per group), and j insulin signaling ( n = 4 per group) were measured in control and M-LRP2 −/− mice. Serum and muscle ApoJ level, body weight, and random blood glucose were measured at 16 weeks of age. Serum insulin levels were measured from overnight fasted mice at 24 weeks of age. ITT was performed at 24 weeks of age and GTT was performed at 18 weeks of age. GSIS was performed at 22 weeks of age. Insulin signaling were performed at 24 weeks of age. All bars and errors represent means ± SEM. p values by two-sided Student’s t -test in a , b , and f , and by repeated measures two-way ANOVA in g and h are indicated. p values for AUCs were evaluated by one-sided Student’s t -test for g and h . p values for AUCs were evaluated by one-sided Student’s t -test for g and h . p values for interaction in g and h were obtained by Greenhouse–Geisser correction. * p < 0.05 vs control by repeated measures two-way ANOVA.

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Quantitation Assay, Control, SDS Page

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: a Insulin-stimulated glucose uptake in muscle, adipose tissue, and brown adipose tissue (BAT) was measured in control and L-ApoJ −/− mice (muscle: n = 20 per group, adipose tissue: n = 16 per group, BAT: n = 20 for control, n = 19 for L-ApoJ −/− ). Mice were studied at 18 weeks of age. b Insulin-stimulated glucose uptake in muscle, adipose tissue, and BAT was measured in control M-LRP2 −/− mice (muscle: n = 10 for control, n = 8 for M-LRP2 −/− , adipose tissue: n = 10 for control, n = 7 for M-LRP2 −/− , BAT: n = 10 for control, n = 8 for M-LRP2 −/− ). Mice were studied at 17 weeks of age. Mice were injected with insulin in combination with [ 14 C]2-deoxyglucose. In vivo 2-deoxyglucose uptake was measured. All bars and errors represent means ± SEM. p values by two-sided Student’s t -test in a and b are indicated.

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: Control, Injection, In Vivo

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: a Serum ApoJ levels in normal cycling (normal) and PCOS subjects. Immunoblots represent serum ApoJ and Apo B48 protein in normal and PCOS subjects. Ponceau S staining indicates that equal amounts of protein in each lane was loaded on the gel. Scatter diagram shows serum ApoJ levels measured by ELISA from normal ( n = 5) and PCOS subjects ( n = 19). b Changes in HOMA2-IR after placebo ( n = 8) or pioglitazone treatment ( n = 10) in PCOS subjects. c Changes in glucose disposal rate after placebo ( n = 8) or pioglitazone ( n = 9) treatment in PCOS subjects. d Changes in serum ApoJ levels after placebo ( n = 7) or pioglitazone treatment ( n = 10) in PCOS subjects. All bars and errors represent means ± SEM. e Relationship of serum ApoJ with fasting insulin levels ( n = 38), HOMA2-IR ( n = 38), HOMA-β (%) ( n = 38), GDR ( n = 40), HDL cholesterol ( n = 31), LDL cholesterol ( n = 31), free fatty acid ( n = 31), and adiponectin ( n = 40) levels in normal and PCOS subjects. p values were obtained by Spearman’s rank correlation analysis and r values indicate Spearman’s correlation coefficient. f Muscle ApoJ levels in normal ( n = 8) and PCOS subjects ( n = 21). PCOS subjects were treated with placebo or pioglitazone. g Muscle LRP2 levels in normal ( n = 6) and PCOS subjects ( n = 19). h Changes in muscle ApoJ levels after placebo ( n = 6) or pioglitazone treatment ( n = 12) in PCOS subjects. i Changes in muscle LRP2 levels after placebo ( n = 6) or pioglitazone ( n = 6) treatment in PCOS subjects. All bars and errors represent means ± SEM. p values by two-sided Student’s t -test in a , c , d , h , and i are indicated.

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: Western Blot, Staining, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Apolipoprotein J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity

doi: 10.1038/s41467-020-15963-w

Figure Lengend Snippet: Circulating ApoJ is dominantly produced by the liver. Liver-derived ApoJ then transports to skeletal muscle where it binds to its receptor LRP2 on the cell surface, leading to increased molecular interaction between LRP2 and IR. The complexes of LRP2 with the IR then undergo co-endocytosis, which is an essential step of insulin signaling in skeletal muscle. As a result, downstream signaling components activate, leading to enhanced glucose transport system activity. Thus, the ApoJ → LRP2 signaling pathway is central in regulation of glucose homeostasis and insulin signaling.

Article Snippet: Animals bearing a LoxP-flanked ApoJ allele (ApoJ loxP/loxP mice) were generated by inGenious Targeting Laboratory (Stony Brook, NY).

Techniques: Produced, Derivative Assay, Activity Assay