1. Figure 1 Lack of endogenous estrogens increases BW and VF in mice fed normal chow or HFD. A, OVX mice (open squares) gain more weight than ovarian-intact mice (sham-operated control group, open circles) from week 9 after ovariectomy when maintained on a chow diet. OVX mice (black squares) gain weight faster than ovarian-intact mice (black circles) from the second week on a HFD. B, OVX mice fed HFD (OVX/HFD) (black bar) accumulate more fat than ovarian-intact mice (S/HFD) (stripped bar) after 12 weeks on this diet. There was no significant difference in the fat mass of OVX mice on chow diet (OVX/chow) (dotted bar) in comparison with ovarian-intact mice on this diet (S/chow) (white bar). C, VF content in OVX mice fed HFD (OVX/HFD) (black bar) was significantly higher than the one present in mice fed a chow diet and in ovarian-intact mice fed HFD. D, Stronger HFD effect regarding the visceral to sc fat accumulation ratio in comparison with ovariectomy effect on this measurement (n = 12–13 per group, all animals were vehicle treated). Results are presented as mean ± SEM (n = 6); ***, P < .001.
From: Estradiol Prevents Fat Accumulation and Overcomes Leptin Resistance in Female High-Fat Diet Mice
Endocrinology. 2014;155(11): doi: /en
Endocrinology | Copyright © 2014 by the Endocrine Society

2. Figure 2 E2 administration reduces the development of obesity in ovarian-intact female mice fed a HFD. A, Mice fed a HFD and on a 12-week cyclic regime of 2 μg/mice of E2 (gray circles) show a lower weight gain compared with HFD vehicle control animals (black circles) and a nonsignificant increased weight gain to chow-fed controls (open circle). B, E2-treated mice fed a HFD (S/E2/HFD) (gray bar) have an increased lean BW compared with mice treated with vehicle fed a regular diet (S/V/chow) (white bar) or HFD (S/V/HFD) (black bar). C, E2-treated mice fed a HFD (S/E2/HFD) (gray bar) have an increased bone mass density (BMD) compared with mice treated with vehicle fed a regular diet (S/V/chow) (white bar) or HFD (S/V/HFD) (black bar). D, E2 treatment (S/E2/HFD) (gray bar) prevents the increment of total fat; (E) VF content and (F) ratio of visceral to sc fat to levels of those found in mice in a regular diet (S/V/chow) (white bar) in contrast to mice fed a HFD (S/V/HFD) (black bar) (n = 6 per group). Results are presented as mean ± SEM (all animals were sham operated). *, P < .05; **, P < .01; ***, P < .001.
From: Estradiol Prevents Fat Accumulation and Overcomes Leptin Resistance in Female High-Fat Diet Mice
Endocrinology. 2014;155(11): doi: /en
Endocrinology | Copyright © 2014 by the Endocrine Society

3. Figure 3 E2 administration reduces FI but does not increase EE and LA in ovarian-intact female mice fed a HFD. A, E2-treated mice (2 μg/mice) fed a HFD (S/E2/HFD) (gray circle) exhibit lower calories intake compared with HFD vehicle control (S/V/HFD) (black circle) and a similar caloric intake as chow fed controls mice (S/V/chow) (open circle). B, iBAT temperature recorded during 72 hours was similar after E2 injection (gray line) and vehicle (black line) in mice fed HFD. Boxed area represents the dark period. C, iBAT UCP1 protein expression was not different in tissue extracts from mice treated with E2 and vehicle. D, Western blot quantification of UCP1 in E2-treated mice (S/E2/HFD) (gray bar) and vehicle-treated mice (S/V/HFD) (black bar). E, LA was not different after E2 injection (gray line) and vehicle (black line) in mice fed HFD. Boxed area represents the dark period. F and G, CLAMS recording of EE were found not different between vehicle (black circles and bars) and E2-treated (gray circles and bars) obese animals. Results are presented as mean ± SEM (n = 6); ***, P < .001.
From: Estradiol Prevents Fat Accumulation and Overcomes Leptin Resistance in Female High-Fat Diet Mice
Endocrinology. 2014;155(11): doi: /en
Endocrinology | Copyright © 2014 by the Endocrine Society

4. Figure 4 E2 administration reverts the relative hypoestrogenism and the disrupted estrous cyclicity provoked by HFD consumption. A, Estrous cycle illustrated as the percentage of days spent in estrus or diestrus in mice treated with vehicle fed a regular diet (S/V/chow) (white bar). HFD consumption disrupted significantly the estrous cyclicity (S/V/HFD) (black bar). B, Twelve weeks of cyclic regime of 2 μg/mice of E2 raised E2 plasma levels (S/E2/HFD) (gray bar) compared with mice treated with vehicle fed a regular diet (S/V/chow) (white bar) or HFD (S/V/HFD) (black bar). Blood samples were obtained 24 hours after sc E2 or vehicle injection. C, Average uterus weight value was significantly higher in E2-treated mice fed a HFD (S/E2/HFD) (gray bar) after 12 weeks of E2 treatment than vehicle-control mice on a regular diet (S/V/chow) or HFD (S/V/HFD). Uterus weight was lower in HFD (S/V/HFD) compared with mice on chow diet (S/V/chow). Results are presented as mean ± SEM (n = 6); *, P < .05; ***, P < .001.
From: Estradiol Prevents Fat Accumulation and Overcomes Leptin Resistance in Female High-Fat Diet Mice
Endocrinology. 2014;155(11): doi: /en
Endocrinology | Copyright © 2014 by the Endocrine Society

5. Figure 5 E2 treatment decreases obesity, fat mass, and FI in obese mice fed HFD. A, OVX obese mice fed HFD (OVX/E2) (gray circles) lost weight during a 4-week cyclic regime of 2 μg/mice of E2 compared with OVX obese mice treated with vehicle (OVX/V) (black circles). A PBW group was also analyzed (mice received HFD to match the BW of the E2-treated group) (OVX/PBW) (white circles). Total fat mass (B) and VF (C) decreased in OVX/E2 and OVX/PBW mice after 4 weeks of treatment compared with their initial values. D, The visceral/sc fat ratio was not modified in any of the 3 groups studied. E, OVX obese mice fed HFD (OVX/E2) (gray circle) display lower cumulative FI during E2 treatment than OVX obese mice treated with vehicle (OVX/V) (black circle) but higher cumulative FI than OVX/PBW mice. Results are presented as mean ± SEM (n = 6); *, P < .05; **, P < .01; ***, P < .001.
From: Estradiol Prevents Fat Accumulation and Overcomes Leptin Resistance in Female High-Fat Diet Mice
Endocrinology. 2014;155(11): doi: /en
Endocrinology | Copyright © 2014 by the Endocrine Society

6. Figure 6 E2 treatment increases EE, LA, and RER in obese mice fed HFD
Figure 6 E2 treatment increases EE, LA, and RER in obese mice fed HFD. A, EE during 24 hours measured by indirect calorimetry in 3 groups of mice: E2-treated OVX obese mice fed HFD (OVX/E2) (gray circles), OVX obese mice treated with vehicle (OVX/V) (black circles), and PBW mice fed HFD (OVX/PBW) (white circles). B, Average of EE during the light and dark periods shows that OVX/E2 mice (gray bar) increase their EE compared with OVX/V (black bar) and OVX/PBW mice (white bar). C, LA during 24 hours measured by indirect calorimetry in each group. D, Average of LA during the light and dark periods shows that OVX/E2 mice (gray bar) increase the EE compared with OVX/V (black bar) and OVX/PBW (white bar) mice during the dark period. E, RER during 24 hours measured by indirect calorimetry. F, Average of RER shows that OVX/E2 mice (gray bar) decrease the RER compared with OVX/V (black bar) and OVX/PBW (white bar) mice during the light and dark period. Gray area represents the dark period. G, iBAT thermogenic marker expression analysis by real-time PCR shows a relative increase of UCP1 signal in OVX/E2 treated (black bars) compared with vehicle control mice (OVX/V/HFD) (white bar). Results are presented as mean ± SEM (n = 6); *, P < .05; ***, P < .001.
From: Estradiol Prevents Fat Accumulation and Overcomes Leptin Resistance in Female High-Fat Diet Mice
Endocrinology. 2014;155(11): doi: /en
Endocrinology | Copyright © 2014 by the Endocrine Society

7. Figure 7 E2 treatment restores leptin sensitivity in obese mice fed HFD. OVX obese mice fed HFD treated with E2 (OVX/E2) lost weight (A) and decrease FI (B) during 48 hours after a single injection of leptin (0.2 μg/mouse) (black bar) into the lateral ventricle compared with aCSF (white bar) injection. Obese mice treated with vehicle (OVX/V) or OVX/PBW mice did not change their BW or FI after leptin treatment (black bar) compared with aCSF injection (white bar). C, Representative microphotographs of hypothalamic sections from OVX/V, OVX/E2, and OVX/PBW mice showing p-STAT-positive staining after 30-minute icv leptin (0.2 μg/μL) or aCSF injection. D, p-STAT3-positive cell number in arcuate nucleus of OVX/E2 after leptin injection (black bar) was higher than after aCSF (white bar). There were not detectable changes in p-STAT3-positive cells on ARH of OVX/V and OVX/PBW mice after leptin injection. E–G, A 45-minute incubation with E2 (10nM) (black bar) increase the α-MSH secretion (E) and decrease the AgRP (F) and NPY (G) secretion in hypothalamic ex vivo slices of mice fed chow and DIO mice leptin resistant. E2-treated brain data (black bar) was normalized to aCSF-treated brain data (white bar) from mice fed chow or HFD. Results are presented as mean ± SEM (n = 6–8); *, P < .05; **, P < .01; ***, P < .001.
From: Estradiol Prevents Fat Accumulation and Overcomes Leptin Resistance in Female High-Fat Diet Mice
Endocrinology. 2014;155(11): doi: /en
Endocrinology | Copyright © 2014 by the Endocrine Society