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1.
Figure 7

Figure 7. From: IRS2 signaling in LepR-b neurons suppresses FoxO1 to control energy balance independently of leptin action.

Levels of mRNA by semi-quantitative RT-PCR of Agrp (A) and Pomc (B) from hypothalamus of 16-week-old chow-fed male mice of the indicated genotypes (n=6). Data are presented as mean ± SEM; *, p < 0.05.

Marianna Sadagurski, et al. Cell Metab. ;15(5):703-712.
2.
Figure 3

Figure 3. From: IRS2 signaling in LepR-b neurons suppresses FoxO1 to control energy balance independently of leptin action.

Glucose homeostasis in LeprΔIrs2 mice. (A) Fasting serum insulin levels (n = 10/genotype) for 6 week-old male mice and (B) 1-year-old mice (n=5/genotype) of the indicated genotypes. Glucose tolerance test of (C) 6-week-old male mice or (D) 24-week-old mice of the indicated genotypes (n=10/genotype). (E) Insulin tolerance test of 7-8 weeks old mice (n=9-10/genotype) or (F) 24-week-old mice (n=8-10/genotype). Data are expressed as mean ± SEM. Irs2L/L -mice versus LeprΔIrs2-mice: *, p < 0.05; **, p < 0.01. See also Figures S3, S4 and Table S1.

Marianna Sadagurski, et al. Cell Metab. ;15(5):703-712.
3.
Figure 5

Figure 5. From: IRS2 signaling in LepR-b neurons suppresses FoxO1 to control energy balance independently of leptin action.

FoxO1 signaling in LeprΔIrs2 –mice. (A) Immunofluorescence for FoxO1 (red) in 6-weeks-old control LeprEGFP and LeprEGFPΔIrs2 mice injected icv with vehicle or insulin (300 mU; 1 hour). Representative images from the hypothalamus of LeprEGFP and LeprEGFPΔIrs2 mice are shown. Scale bar: 100 μm. Lower (small) panels are digital zooms of boxed areas from the top panel, showing FoxO1 (red) alone (left panels) or merged with and EGFP (green) (right panels) in 6-week-old mice of the indicated genotypes. (B) Quantification of LepR-b/EGFP neurons containing nuclear FoxO1 immunoreactivity (n=4/genotype). Data are presented as mean ± SEM; *, p < 0.05.

Marianna Sadagurski, et al. Cell Metab. ;15(5):703-712.
4.
Figure 6

Figure 6. From: IRS2 signaling in LepR-b neurons suppresses FoxO1 to control energy balance independently of leptin action.

Restoration of energy homeostasis in LeprΔIrs2 mice by deletion of FoxO1 from LepR-b neurons. Parameters of energy balance for 16-week-old chow-fed male mice of the indicated genotypes (n=10/genotype): (A) Body weight, (B) food intake. Parameters of energy balance for 16-week-old chow-fed male mice of the indicated genotypes: (C) oxygen consumption (O2, L•kg-1•h-1), and (D) carbon dioxide production (CO2, L•kg-1•h-1) during the dark cycle. (E) Glucose tolerance test and (F) fasting serum insulin concentrations in 16-week-old mice of the indicated genotypes (n=8-10/genotype). Data are presented as mean ± SEM; *, p < 0.05. See also Figure S6 and S7.

Marianna Sadagurski, et al. Cell Metab. ;15(5):703-712.
5.
Figure 4

Figure 4. From: IRS2 signaling in LepR-b neurons suppresses FoxO1 to control energy balance independently of leptin action.

LepR-b signaling and leptin sensitivity in LeprΔIrs2 mice. (A) Immunostaining for pSTAT3 in 6-week-old mice of the indicated genotypes injected intraperitoneally with vehicle or leptin (5 mg/kg, i.p., 2 hours, 3v- 3rd cerebral ventricle). Representative images from the hypothalamus are shown. Scale bar: 100 μm (B) Quantification of pSTAT3 immunoreactivity (n=4/genotype). (C and D) Cumulative food intake and reduction in body weight were measured over a 3-day period of treatment with leptin (5 mg/kg i.p.; BID) or vehicle in 6-week-old male mice of the indicated genotypes. Data are presented as mean ± SEM; *, p < 0.05; **, p < 0.01. See also Figure S5.

Marianna Sadagurski, et al. Cell Metab. ;15(5):703-712.
6.
Figure 1

Figure 1. From: IRS2 signaling in LepR-b neurons suppresses FoxO1 to control energy balance independently of leptin action.

LeprΔIrs2 mice are obese. (A) Schematic representation of our breeding strategy to generate LeprΔIrs2 mice. (B) Average body weights of male LeprΔIrs2 -mice (closed circles) and control Irs2L/L mice (open circles) on regular chow diet was determined in each average age group by generalized linear regression (SPSS, v19). The number of mice in each group is indicated in parentheses (mean ± SD; *, Bonferroni p<0.001). (C) Percent body fat and (D) lean body mass was determined by DEXA using 12-week-old Irs2L/L (n=12) and LeprΔIrs2 (n=12) mice (mean ± SEM; *, p<0.05). (E) Serum leptin levels of 12-week-old male Irs2L/L (n=7) and LeprΔIrs2 (n=7) mice (mean ± SEM; *, p<0.05). (F) Food intake over 48 hours in 12-week-old male Irs2L/L (n=12) and LeprΔIrs2 (n=12) mice fed regular chow diet (mean ± SEM; *, p<0.05). See also Figure S1.

Marianna Sadagurski, et al. Cell Metab. ;15(5):703-712.
7.
Figure 2

Figure 2. From: IRS2 signaling in LepR-b neurons suppresses FoxO1 to control energy balance independently of leptin action.

Energy expenditure in LeprΔIrs2 mice. 24 week-old male mice of the indicated genotype were monitored for 72 hours in the CLAMS (n=10/genotype) to assess (A) oxygen consumption (O2, l/kg/h), (B) carbon dioxide production (CO2, l/kg/h), (C) heat production (kcal/kg 0.75) and (D) locomotor activity during the light and dark cycles. Each group was analyzed by generalized linear regression (SPSS, v19), (mean ± SD; *, Bonferroni p<0.001). (E) Representative H&E staining of brown adipose tissue (BAT) of Irs2L/L (left panel) and LeprΔIrs2 mice (right panel) mice aged 24 weeks. Scale bar: 500 μm. (F) Levels of mRNA by semi-quantitative RT-PCR of Ucp1 and Ppargc1α from BAT of 24-week-old Irs2L/L and LeprΔIrs2 mice (n=6). Data are presented as mean ± SEM; *, p < 0.05. See also Figure S2.

Marianna Sadagurski, et al. Cell Metab. ;15(5):703-712.

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