IMH photographs of VPAC2-R protein expression.

<p>(A) Section of a human ovary from the same fetus as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037015#pone-0037015-g001" target="_blank">Figure 1</a> (A) and (D). Note the red-brown staining indicating VPAC2-R expression in the oocytes (full weak cytoplasmic staining with nuclear staining), in a portion of the GC and stroma cells. Original magnification X400. (B) Section of human ovary from the same woman as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037015#pone-0037015-g002" target="_blank">Figure 2</a> (A) and (B). Note the secondary follicle (full cytoplasmic staining with nuclear staining), with red-brown staining in the GC and stroma cells. Original magnification X400. (C) Negative control for the same ovarian section as in panel A. Note the primordial follicles with overall blue staining and lack of red-brown staining. Original magnification X400. (D) Negative control for the same ovarian section as in panel B. Note the primordial follicles with overall blue staining and lack of red-brown staining. Original magnification X400.</p

Protein expression (IMH) of VIP, VPAC1-R and VPAC2-R in human ovaries.

<p><u>Note</u>: IMH = immunohistochemistry, GW = gestational weeks. Percentages represent the proportion of the samples with follicular staining.</p><p>Staining intensities: + = positive staining; full/partial = full or partial cytoplasmic staining.</p

IMH photographs of VIP protein expression.

<p>(A) Section of human ovary from a 22-GW-old fetus. Note the primordial follicles, weak red-brown staining indicating VIP expression in oocytes (full cytoplasmic and nuclear staining), and weak staining in a portion of the GC and stroma cells. Original magnification X400. (B) Section of human ovary from a 6-year-old girl. Note the primordial follicles with red-brown staining indicating VIP protein expression in the oocyte (mainly cytoplasmic staining and nuclear staining), and in a portion of the GC and a portion of the stroma cells. Original magnification X400. (C) Positive control for VIP protein expression of section of mouse brain. Note the red-brown staining in the sample. Original magnification X400. (D) Negative control for the same ovarian section as in panel A. Note the primordial follicles with the overall blue staining and the lack of red-brown staining. Original magnification X400. (E) Negative control for the same ovarian section as in panel B. Note the primordial follicles with overall blue staining and the lack of red-brown staining. Original magnification X400.</p

Representative RT-PCR gel illustrating expression of the VIP, VPAC1-R and VPAC2-R genes in fetal and adult ovaries.

<p>(A) VIP gene (134 base pairs), (B) VPAC1-R gene (97 base pairs), (C) VPAC2-R gene (155 base pairs), (D) DHFR gene as positive control (231base pairs). Sample 1: Ovary from a 21-year-old woman, Sample 2: Ovary from a 29-year-old woman, Sample 3: Ovary from a 39-year-old woman, Sample 4: Ovary from a 21-GW-old fetus, Sample 5: Ovary from a 23-GW-old fetus, Sample 6: Ovary from a 25-GW-old fetus, Sample 7: Ovary from a 27-GW-old fetus. M: Marker (100 base pair DNA ladder, QIAGEN), +: RT presence, -: RT absence.</p

IMH photographs of VPAC1-R protein expression.

<p>(A) Section of human ovary from a 22-year-old woman. Note the primordial follicles, with red-brown staining indicating VPAC1-R expression in the oocytes (full cytoplasmic staining and nuclear staining), and in a portion of the GC and stroma cells. Original magnification X400. (B) Negative control for the same ovarian section as in panel A. Note the primordial follicle, overall blue staining, and lack of red-brown staining. Original magnification X400.</p