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2.
Figure 2

Figure 2. From: Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation.

The AR is expressed in Xenopus oocytes. Western blots were performed on extracts of oocytes injected with buffer (mock) or cRNA encoding the Xenopus AR (XAR).

L. B. Lutz, et al. Proc Natl Acad Sci U S A. 2001 November 20;98(24):13728-13733.
3.
Figure 5

Figure 5. From: Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation.

Metabolism of and responses to DHEA in oocytes with and without 3βHSDII. (A) DHEA (circles) and AD- (squares) induced maturation of oocytes injected with either buffer (mock, closed symbols) of cRNA encoding 3βHSDII (HSDII, open symbols) was measured (n = 20 oocytes). (B) DHEA metabolism in the injected oocytes was examined 48 h after injection by incubating oocytes in MBSH containing 100 nM radiolabeled DHEA for 4 h. Steroids were extracted from the media or oocytes and resolved by TLC (30). The remaining spots represent <10% of the total counts. Each of these experiments was performed twice with equivalent results.

L. B. Lutz, et al. Proc Natl Acad Sci U S A. 2001 November 20;98(24):13728-13733.
4.
Figure 3

Figure 3. From: Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation.

Flutamide inhibition of androgen-induced maturation and p42 phosphorylation. Oocytes were incubated in MBSH in the presence or absence of flutamide with deoxycorticosterone (DOC) (A), progesterone (B), testosterone (C), or AD (D). Similar experiments were performed 3, 5, 5, and 3 times, respectively, with nearly identical results. (E) Ooocytes were treated as above except incubated with the indicated steroid at 50 nM for 4 h. Blots were probed with antiserum against phosphorylated p42 protein (Upper) followed by antiserum against total p42 protein (Lower). Twenty oocytes were used for each sample, and similar experiments were performed three times with equivalent results.

L. B. Lutz, et al. Proc Natl Acad Sci U S A. 2001 November 20;98(24):13728-13733.
5.
Figure 1

Figure 1. From: Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation.

Steroid production by β-hCG-stimulated ovaries and their effects on oocyte maturation. In vivo steroid levels were determined after injection of frogs with 500 units of β-hCG. (A) Steroids were extracted from ovaries at the indicated times, and RIAs were performed. Steroid levels are displayed as nanograms of steroid per gram of tissue ± SEM (n = 3). (B) Serum steroid concentrations 8 h after β-hCG injection of three β-hCG-injected frogs are displayed as nM ± SEM (n = 2). (C) In vitro ovarian steroid production was measured by incubating pieces of ovary in MBSH with 10 units of β-hCG. Buffer was removed at the indicated times and steroid concentrations determined. Steroid production is displayed as nanograms of steroid per gram of ovarian tissue ± SEM (n = 2). (D) Maturation responses of oocytes to androgens and progesterone were determined (n = 20 oocytes). (E) Progesterone (P), AD (A), and testosterone (T) at 500 nM promote dephosphorylation of cdc2 at 8 h relative to cells treated with ethanol alone (−) (Upper). Total cdc2 is equal in all samples (Lower). All experiments were performed at least three times with nearly identical results.

L. B. Lutz, et al. Proc Natl Acad Sci U S A. 2001 November 20;98(24):13728-13733.
6.
Figure 4

Figure 4. From: Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation.

Progesterone metabolism by isolated Xenopus oocytes and its effects on maturation. (A) Progesterone metabolism was examined by incubating 40 oocytes for the times indicated with 10 nM (lane 8) or 100 nM (lanes 1–7) radiolabeled progesterone. Oocytes in the lower panel were preincubated with ketoconazole followed by addition of 100 nM radiolabeled progesterone. Steroids were extracted from the media or oocytes and examined by TLC. Locations of progesterone (Prog), AD (Andro), and 17OHP are indicated. The identities of these steroids were confirmed by HPLC. The other minor components were <5% of the total counts. (B) Oocytes (250) were injected with radiolabeled progesterone and incubated in MBSH for 4 h. Steroids were extracted from the oocytes (O) or medium (M) and treated as above. Lane 1 is radiolabeled progesterone (0). (C) Autoradiograms of progesterone metabolites in COS cells transfected with pcDNA3.1 (M) or cDNA encoding Xenopus CYP17 (XC17) or Human CYP17 (HC17). (D) Maturation responses to progesterone (Prog, circles) and AD (Andro, squares) in the presence (open symbols) or absence (closed symbols) of ketoconazole (n = 20 oocytes). Each experiment was performed at least three times with equivalent results.

L. B. Lutz, et al. Proc Natl Acad Sci U S A. 2001 November 20;98(24):13728-13733.

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