Journal: Endocrinology

Article Title: Opposite effects of androgens and estrogens on adipogenesis in rat preadipocytes: evidence for sex and site-related specificities and possible involvement of insulin-like growth factor 1 receptor and peroxisome proliferator-activated receptor gamma2.

doi: 10.1210/endo.141.2.7293

Figure Lengend Snippet: FIG. 1. Effect of 17b-estradiol on the cell growth of sc and parametrial preadipocytes from ovariectomized female rats. After 3 days of culture in DMEM with 2% FCS, in the presence or absence of 17b-estradiol (0.01 mM) or in the presence of 17b-estradiol (0.01 mM) and ICI182780 (1 mM) simultaneously, cells were collected and counted. Data are the means 6 SEM of six to eight separate experiments. Each experiment was performed in triplicate. *, P , 0.05; CONT, control; EST, 17b- estradiol; ICI, ICI182780.

Article Snippet: DMEM and DMEM-F12 (phenol red free), HEPES, porcine insulin, transferrine, T3, NADH, H1, dihydroxyacetone phosphate, testosterone, 5a-dihydrotestosterone (DHT), 17b-estradiol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate, estrone (1,3,5 [10]-estratrien 3 ol 17 one), androstanediol (5a- androstane 3b, 17b, diol), androstenediol (5androstene 3b, 17b, diol), and BSA were from Sigma (St. Louis, MO); ICI182780 was from Tocris (Bristol, UK); RU23908 was from Roussel-Uclaf (Romainville, France); collagenase was from Roche Molecular Biochemicals (Mannheim, Germany); FCS was from Life Technologies (Grand Island, NY); polyclonal rabbit anti-PPARg2 antiserum was from Affinity BioReagents, Inc. (Golden, CO); polyclonal rabbit anti-IGF1 receptor (anti-IGF1R) antiserum and polyclonal goat anti IGF1 antiserum were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA); and horseradish peroxidase-labeled rabbit anti-IgG was from Sanofi Pharmaceuticals, Inc. Pasteur (Marne la Coquette, France).

Techniques: Control

Journal: Endocrinology

Article Title: Opposite effects of androgens and estrogens on adipogenesis in rat preadipocytes: evidence for sex and site-related specificities and possible involvement of insulin-like growth factor 1 receptor and peroxisome proliferator-activated receptor gamma2.

doi: 10.1210/endo.141.2.7293

Figure Lengend Snippet: FIG. 2. Effect of various sex steroids on GPDH activity of epididymal, parametrial, and femoral sc preadipocytes from male and ovariectomized female rats. Precursor cells from epididymal, parametrial, and sc fat deposits were cultured as described in Materials and Methods. Testosterone, DHT, and RU23908 (Fig. 2, A and B), 17b-estradiol, and ICI182780 (Fig. 2, C and D) at 0.1 mM, 10 mM, or 100 mM were added during all the differentiation process in ITT medium. After 7 to 8 days in ITT medium, cells were collected, and GPDH activity was measured. Values are expressed as percentage of the mean values obtained for cells maintained in culture without steroids (1886 6 220 mU/mg protein and 928 6 106 mU/mg protein in epididymal and sc cells from male rats, respectively, and 1175 6 275 mU/mg protein and 2545 6 750 mU/mg protein in parametrial and sc cells from ovariectomized female rats, respectively). Each value represents the mean 6 SEM of four to seven separate experiments. *, P , 0.05; **, P , 0.01; ns, not significant; (a), vs. control; (b), vs. RU23908 in Fig. 2A; (b), vs. ICI182780 in Fig. 2D.

Article Snippet: DMEM and DMEM-F12 (phenol red free), HEPES, porcine insulin, transferrine, T3, NADH, H1, dihydroxyacetone phosphate, testosterone, 5a-dihydrotestosterone (DHT), 17b-estradiol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate, estrone (1,3,5 [10]-estratrien 3 ol 17 one), androstanediol (5a- androstane 3b, 17b, diol), androstenediol (5androstene 3b, 17b, diol), and BSA were from Sigma (St. Louis, MO); ICI182780 was from Tocris (Bristol, UK); RU23908 was from Roussel-Uclaf (Romainville, France); collagenase was from Roche Molecular Biochemicals (Mannheim, Germany); FCS was from Life Technologies (Grand Island, NY); polyclonal rabbit anti-PPARg2 antiserum was from Affinity BioReagents, Inc. (Golden, CO); polyclonal rabbit anti-IGF1 receptor (anti-IGF1R) antiserum and polyclonal goat anti IGF1 antiserum were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA); and horseradish peroxidase-labeled rabbit anti-IgG was from Sanofi Pharmaceuticals, Inc. Pasteur (Marne la Coquette, France).

Techniques: Activity Assay, Cell Culture, Control

Journal: Endocrinology

Article Title: Opposite effects of androgens and estrogens on adipogenesis in rat preadipocytes: evidence for sex and site-related specificities and possible involvement of insulin-like growth factor 1 receptor and peroxisome proliferator-activated receptor gamma2.

doi: 10.1210/endo.141.2.7293

Figure Lengend Snippet: FIG. 5. Effect of estrogens on IGF1R expression in sc preadipocytes from ovariectomized female rats. sc precursor cells were cultured as described in Materials and Methods. 17b-estradiol and ICI182780 were added during 4 days after plating. Preadipocytes were collected, and membrane extracts were prepared as described in Materials and Methods. A, Western-blot analysis from one representative experiment. Forty micrograms of protein were immunoblotted with polyclonal rabbit anti-IGF1R antiserum; B, Densitometric analysis [values are mean 6 SEM obtained from three to four separate experiments and are expressed as control values (without steroids)]. **, P , 0.05; (a), vs. control; (b), vs. ICI182780.

Article Snippet: DMEM and DMEM-F12 (phenol red free), HEPES, porcine insulin, transferrine, T3, NADH, H1, dihydroxyacetone phosphate, testosterone, 5a-dihydrotestosterone (DHT), 17b-estradiol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate, estrone (1,3,5 [10]-estratrien 3 ol 17 one), androstanediol (5a- androstane 3b, 17b, diol), androstenediol (5androstene 3b, 17b, diol), and BSA were from Sigma (St. Louis, MO); ICI182780 was from Tocris (Bristol, UK); RU23908 was from Roussel-Uclaf (Romainville, France); collagenase was from Roche Molecular Biochemicals (Mannheim, Germany); FCS was from Life Technologies (Grand Island, NY); polyclonal rabbit anti-PPARg2 antiserum was from Affinity BioReagents, Inc. (Golden, CO); polyclonal rabbit anti-IGF1 receptor (anti-IGF1R) antiserum and polyclonal goat anti IGF1 antiserum were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA); and horseradish peroxidase-labeled rabbit anti-IgG was from Sanofi Pharmaceuticals, Inc. Pasteur (Marne la Coquette, France).

Techniques: Expressing, Cell Culture, Membrane, Western Blot, Control

Journal: Oncogene

Article Title: High estrogen receptor alpha activation confers resistance to estrogen deprivation and is required for therapeutic response to estrogen in breast cancer

doi: 10.1038/s41388-021-01782-w

Figure Lengend Snippet: (A) Cells were seeded at low density and treated in triplicate as indicated for 4 wk. Colonies were fixed and stained with crystal violet, and colony area was quantified. Parental cells were hormone-deprived (HD) for 3 d prior to seeding. (B) Lysates from parental cells (hormone-deprived for 3 d) and LTED cells were analyzed by immunoblot. (C) RNA isolated from parental cells (hormone-deprived for 3 d) and LTED cells was analyzed by RT-qPCR. Values for indicated transcripts were normalized to ACTB expression. (D) LTED cells were seeded at low density and treated in triplicate ± fulvestrant (fulv) as indicated for 4 wk. Colony area was analyzed as in (A). (E) LTED cells were treated ± 1 μM fulv for 3 d. RNA was isolated and analyzed by RT-qPCR as in (B). (F) Ovx mice bearing WHIM16 tumors were randomized to treatment as indicated. (G) Tumors were harvested from mice treated as in (F) after 8 d of treatment. Expression of ER, Ki67, cleaved caspase 3 (CC3) and progesterone receptor (PR) were analyzed by IHC. *p≤0.05, **p≤0.005, ***p≤0.0005 by two-tailed t-test (D, G), Bonferroni multiple comparison-adjusted post-hoc test (C, E), or linear mixed modeling (F). Data are shown as mean ± SD.

Article Snippet: For long-term E2 treatment, pellets were replaced every 30 d. Fulvestrant (Ontario Chemicals, Inc., Guelph, ON, Canada) was dissolved in ethanol at 500 mg/mL, and diluted 10-fold with castor oil to yield a 50-mg/mL solution.

Techniques: Staining, Western Blot, Isolation, Quantitative RT-PCR, Expressing, Paraffin-embedded Immunohistochemistry, Two Tailed Test, Comparison