In vivo studies of 129S6 mice.

(A) Body weight gain during treatment with DEHP (B) Insulin tolerance test of 129S6 mice (N = 8 controls, N = 8 DEHP treatment) after 6 weeks of DEHP intake (C) fat mass and (D) lean mass as percent of body weight was determined in awake female mice by using nuclear magnetic resonance technology with EchoMRI700 instrument (Echo Medical Systems, Houston, TX, USA) at the end of observation period (10 weeks). Data are presented as percentage of total body fat and lean mass from body weight. Results are expressed as means ± SE from at least 5 female animals per treatment group. (E) Food intake per animal, day and body weight calculated from food intake and body weight measurements in weeks 5 and 6 of treatment. (F) ipGTT was performed on 4-h-fasted after 8 weeks of DEHP treatment in female mice. Results are expressed as means ± SE from at least 5 female animals per treatment group. (G) Serum adiponectin and (H) estrogen concentrations were analyzed at the end of observation period (10 weeks) in female mice (n = 5 per treatment group). Data are presented as mean ± SE from at least 5 female animals per treatment group. (I) Western Blot quantification of estrogen receptor protein expression (Esr1), (J) glucocorticoid receptor (Glur), Adiponectin (K) and Ppary (L) in subcutaneous (SC) and visceral adipose tissue of female 129S6 mice (at least n = 3 per experimental group). (M) Representative images in adipose tissue of control and DEHP treated mice. Equal protein loading was verified using mouse anti-D-glyceraldehyde-3-phosphate dehydrogenase (Gapdh) antibody. The different degrees of significance (t-test with Welch correction) were indicated as follows in the graphs. *p<0.05; ** p< 0.01.