recombinant mouse gdf9 protein  (R&D Systems)

Journal: NPJ Microgravity

Article Title: Simulated microgravity reduces quality of ovarian follicles and oocytes by disrupting communications of follicle cells

doi: 10.1038/s41526-023-00248-5

Figure Lengend Snippet: a Illustration of the strategy to label the Oo-Mvi by Gdf9-Cre;mTmG mouse model. The membrane-localized red fluorescent protein (mT) switches to green-fluorescent protein (mG) in oocytes of Gdf9-Cre;mTmG mouse to label oocyte membrane morphology. b Images of Gdf9-Cre;mTmG oocytes, showing the mushroom-like Oo-Mvi with vesicle tips distributed in the zona pellucida of oocytes in both the SMG and the NG groups. Scale bars, 30 μm. c 3D high-resolution images showing a decreased density of Oo-Mvi on the oocytes’ surface under SMG. Scale bars, 10 μm. d Numbers of Oo-Mvi reduced in SMG oocytes ( n = 8) compared to that in NG ( n = 8), showing a significantly reduced number of Oo-Mvi on oocytes in follicles after SMG treatment. p value = 0.00052. e High magnification showing that the length of Oo-Mvi in the SMG group was shorter than that in the NG group. Scale bars, 5 μm. f Quantification of the length of Oo-Mvi confirmed a dramatic decrease in the SMG group ( n = 30) compared to that in the NG group ( n = 30). p value = 0.00000000000000000052. The colors were inverted to black/white (b/w) to highlight Oo-Mvi in ( e ). Representative images are shown. Data is presented as the mean ± SD. Data were analyzed by two-tailed unpaired Student’s t -test and *** P < 0.001.

Article Snippet: In the GDF9-supplying experiment, we added GDF9 protein (500 ng/mL, 739-G9-010/CF, R&D) into follicle cultured medium for 2 days.

Techniques: Two Tailed Test

Journal: NPJ Microgravity

Article Title: Simulated microgravity reduces quality of ovarian follicles and oocytes by disrupting communications of follicle cells

doi: 10.1038/s41526-023-00248-5

Figure Lengend Snippet: a Relative mRNA levels of Gdf9, Bmp15 , and Fgf8 under the SMG or the NG group, showing a decreased expression of OSFs in the SMG group ( n = 4). Gdf9: p value = 0.0081, Bmp15: p value = 0.0032, Fgf8: p value = 0.021. b Relative mRNA levels of Fscn1 and Myo10 under the SMG or the NG group, showing that the expression of GC-TZP forming related genes was downregulated after SMG treatment ( n = 4). Fscn1: p value = 0.025, Myo10: p value = 0.021. c Supplying GDF9 increased the PB1 ratio of oocytes (red arrowheads) from SMG treated follicles. Scale bars, 100 μm. d The ratio of PB1 in different groups, showing that the GDF9 supplement significantly increased the maturation of oocytes ( n = 38 in NG, n = 35 in SMG and n = 61 in SMG + GDF9). NG v.s. SMG: p value = 0.00021, SMG v.s. SMG + GDF9: p value = 0.021. Representative images of oocytes are shown. Data are presented as the mean ± SD. Data were analyzed by two-tailed unpaired Student’s t -test in ( a , b ) and two-way ANOVA in ( d ). n.s. P ≥ 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In the GDF9-supplying experiment, we added GDF9 protein (500 ng/mL, 739-G9-010/CF, R&D) into follicle cultured medium for 2 days.

Techniques: Expressing, Two Tailed Test

Journal: Scientific reports

Article Title: Cooperative effects of oocytes and estrogen on the forkhead box L2 expression in mural granulosa cells in mice.

doi: 10.1038/s41598-022-24680-x

Figure Lengend Snippet: Figure 4. Effects of BMP15 and GDF9 signals on FOXL2 expression in MGCs. (A) MGCs were cultured with or without recombinant BMP15/GDF9 (ODPFs) and E2 for 24 h (n = 4). (B) MGCs cultured with both oocytes and E2 were treated with or without an inhibitor of ALK5, SB431542 for 24 h (n = 4). (C) Comparison of the expression levels of FOXL2 protein between MGCs of Bmp15−/−/Gdf9+/− mice (DM) and those of littermate control Bmp15+/−/Gdf9+/− mice (Ctrl) (n = 3 each). The expression of FOXL2 protein was examined by western blotting (upper panel) and band intensities were quantified (lower panel). Asterisks or different letters (a and b) denote significant differences (p < 0.05).

Article Snippet: In some experiments, the basic culture medium was supplemented with 10–7 M of 17β-estradiol (Sigma-Aldrich), recombinant human BMP15 (50 ng/ml; R&D Systems, Minneapolis, MN, USA), recombinant mouse GDF9 (50 ng/ml; R&D Systems), and/or an inhibitor of ALK5, SB431542 (10 μM; FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan).

Techniques: Expressing, Cell Culture, Recombinant, Comparison, Control, Western Blot