1. Growth differentiation factor 3 is induced by bone morphogenetic protein 6 (BMP-6) and BMP-7 and increases luteinizing hormone receptor messenger RNA expression in human granulosa cells 
Jia Shi, M.D., Ph.D., Osamu Yoshino, M.D., Ph.D., Yutaka Osuga, M.D., Ph.D., Ikumi Akiyama, M.D., Miyuki Harada, M.D., Ph.D., Kaori Koga, M.D., Ph.D., Akihisa Fujimoto, M.D., Ph.D., Tetsu Yano, M.D., Ph.D., Yuji Taketani, M.D., Ph.D. 
Fertility and Sterility 
Volume 97, Issue 4, Pages (April 2012)
DOI: /j.fertnstert
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Immunohistochemistry of growth differentiation factor (GDF)-3 in human ovary. GDF-3 protein localization was examined in normal human ovaries. Ovarian tissues were fixed in neutral-buffered formalin and embedded in paraffin blocks. The sections were stained with 10 μg/mL anti-GDF-3 antibody or rabbit IgG as negative control. GDF-3 antibody, which had been preabsorbed with recombinant GDF-3, was also used as a preabsorption control. Representative data from eight specimens were shown. (A, B) Primordial follicle. (C) Primary follicle. (D) Secondary follicle. (E–I) Antral follicle. A, C, D, E, G: Anti-GDF-3 antibody. B, F, H: Negative control antibody. I: GDF-3 antibody, which had been preabsorbed with recombinant GDF-3, was used as a preabsorption control. E, F: lower magnification, ×40. A–D, G–I: higher magnification, ×200.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Effect of bone morphogenetic protein 6 (BMP-6) and BMP-7 on growth differentiation factor 3 (GDF-3) mRNA expression. Human granulosa cells (GC) were cultured with BMP-6 or BMP-7 for 24 hours. The concentration of BMP cytokines was 100 ng/mL. Total RNA was extracted from the GC and subjected to quantitative real-time polymerase chain reaction (PCR) to determine GDF-3 mRNA levels. Data were normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA levels. Representative data from three different experiments was shown as the mean ± standard deviation (SD) relative to an adjusted value of 1.0 for the mean value of the each control. ∗P<.01 versus control.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Effect of growth differentiation factor 3 (GDF-3) on luteinizing hormone (LH) receptor mRNA expression. Human granulosa cells (GC) were cultured with GDF-3 (60–100 ng/mL) in the presence or absence of bone morphogenetic protein 7 (BMP-7) (10–100 ng/mL) for 24 hours. GDF-3 was added 30 minutes before BMP-7 incubation. Total RNA was extracted from the GC and subjected to quantitative real-time polymerase chain reaction (PCR) to determine the LH receptor mRNA levels. Data were normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA levels. Representative data from three different experiments was shown as the mean ± standard deviation (SD) relative to an adjusted value of 1.0 for the mean value of the each control. ∗P<.05 versus control.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions