Journal: Endocrinology

Article Title: Chemerin, a novel regulator of follicular steroidogenesis and its potential involvement in polycystic ovarian syndrome.

doi: 10.1210/en.2012-1424

Figure Lengend Snippet: FIG. 2. Chemerin levels in serum and ovarian cells in CTL and DHT-treated rats. A, Serum (0.5 l) from CTL and DHT-treated rats was subjected to Western blot and immunoblotted with antichemerin antibody. Immunoblots shown are six representative serum samples in each experimental group, whereas each histogram shows results from 12 rats. B, Chemerin and CMKLR1 contents in whole-ovary extracts from CTL and DHT-treated rats were analyzed by Western blot and normalized to GAPDH. Each sample was collected from one ovary of individual CTL or DHT rats (15 rats per histogram), whereas six representative samples were presented in the immunoblots. C, Total RNA were extracted from whole ovaries of CTL and DHT-treated rats. Each sample was collected from one ovary of CTL or DHT rats (nine rats per histogram). The mRNA abundances of chemerin and CMKLR1 were analyzed by real-time PCR and normalized to the expression of GAPDH. D, Granulosa cells were collected from four to five rats of the CTL or DHT group and pooled for total RNA extraction. The mRNA abundances of chemerin and CMKLR1 were analyzed by real-time PCR and normalized to the expression of GAPDH. Data are presented as mean SEM of five independent experiments. Panels A–D used an unpaired Student t test. *, P 0.05, ***, P 0.001 compared with CTL group. NS, Not significant.

Article Snippet: Recombinant mouse active chemerin and antichemerin antibody were from R&D Systems (Minneapolis, MN).

Techniques: Western Blot, Real-time Polymerase Chain Reaction, Expressing, RNA Extraction

Journal: Endocrinology

Article Title: Chemerin, a novel regulator of follicular steroidogenesis and its potential involvement in polycystic ovarian syndrome.

doi: 10.1210/en.2012-1424

Figure Lengend Snippet: FIG. 3. Effect of chemerin on steroidogenic enzyme expression and steroid production in granulosa cells from CTL and DHT-treated rats. Granulosa cells from CTL and DHT-treated rats were cultured with chemerin (0–100 ng/ml) for 48 h, and estradiol and progesterone levels in spent medium were assessed by EIA. Data are presented as mean SEM of three independent experiments and analyzed by two- way ANOVA followed by Bonferroni post hoc test. ***, P 0.001 compared with CTL group; #, P 0.05, ##, P 0.01 compared with untreated control (chemerin 0).

Article Snippet: Recombinant mouse active chemerin and antichemerin antibody were from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Cell Culture, Control

Journal: Endocrinology

Article Title: Chemerin, a novel regulator of follicular steroidogenesis and its potential involvement in polycystic ovarian syndrome.

doi: 10.1210/en.2012-1424

Figure Lengend Snippet: FIG. 4. Effect of chemerin on FSH-induced estradiol secretion and aromatase expression in cultured follicles and granulosa cells. A, Follicles were cultured with chemerin (0–100 ng/ml) FSH (10 ng/ml) for 4 d in vitro, and estradiol levels in spent medium were measured by EIA. Data are presented as mean SEM of 26–30 follicles per group (from three independent experiments). B–D, Granulosa cells were cultured with chemerin (0–100 ng/ml) FSH (100 ng/ml) for 24 h. Testosterone (0.5 M) was added as substrate of aromatase during culture. Estradiol levels in spent medium (B) were measured by EIA. Aromatase mRNA (C) and protein levels (D) were examined by real-time PCR and Western blot, respectively. Data are presented as mean SEM of three to five independent experiments and analyzed by two-way ANOVA followed by Bonferroni post hoc test. *, P 0.05, **, P 0.01, ***, P 0.001 compared with FSH-only group.

Article Snippet: Recombinant mouse active chemerin and antichemerin antibody were from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Cell Culture, In Vitro, Real-time Polymerase Chain Reaction, Western Blot

Journal: Endocrinology

Article Title: Chemerin, a novel regulator of follicular steroidogenesis and its potential involvement in polycystic ovarian syndrome.

doi: 10.1210/en.2012-1424

Figure Lengend Snippet: FIG. 5. Effect of chemerin on FSH-induced progesterone secretion and p450scc expression in cultured follicles and granulosa cells. A, Follicles were cultured with chemerin (0–100 ng/ml) FSH (10 ng/ml) for 4 d in vitro, and progesterone levels in spent medium was examined by EIA. Data are presented as mean SEM of 26–30 follicles per group (from three independent experiments). B–D, Granulosa cells were cultured with chemerin (0–100 ng/ml) FSH (100 ng/ml) for 24 h. Progesterone level in spent medium (B) was measured by EIA. p450scc mRNA (C) and protein levels (D) were examined by real-time PCR and Western blot, respectively. Data are presented as mean SEM of three to five independent experiments and analyzed by two-way ANOVA followed by a Bonferroni post hoc test. *, P 0.05, **, P 0.01, ***, P 0.001 compared with FSH only group.

Article Snippet: Recombinant mouse active chemerin and antichemerin antibody were from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Cell Culture, In Vitro, Real-time Polymerase Chain Reaction, Western Blot

Journal: Endocrinology

Article Title: Chemerin, a novel regulator of follicular steroidogenesis and its potential involvement in polycystic ovarian syndrome.

doi: 10.1210/en.2012-1424

Figure Lengend Snippet: FIG. 6. Effect of chemerin on forskolin- or NR5a1/2-induced aromatase and p450scc expression in granulosa cells. A, Granulosa cells were cultured with chemerin (0–100 ng/ml) forskolin (FSK; 10 M) for 24 h, and the contents of aromatase and p450scc were examined by Western blot. Representative immunoblots and mean SEM of three replicates are shown. Results were analyzed by two-way ANOVA, followed by a Bonferroni post hoc test. **, P 0.01 compared with FSK-only group; ###, P 0.001 compared with CTL group (FSK 0). B and C, Granulosa cells were infected with adenoviral NR5a1B or NR5a2 (C; MOI 10, adenoviral-GFP as control) for 24 h and then cultured with chemerin (0–100 ng/ml, 48 h). The contents of aromatase, p450scc, NR5a1, and NR5a2 were examined by Western blot. Representative immunoblots and mean

Article Snippet: Recombinant mouse active chemerin and antichemerin antibody were from R&D Systems (Minneapolis, MN).

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