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Fig. 1 | Journal of Ovarian Research

Fig. 1

From: Elucidating Decorin’s role in the preovulatory follicle

Fig. 1

a. Expression of DCN in CGCs and MGCs. CGCs and MGCs were aspirated from preovulatory follicles (> 17 mm) of woman undergoing IVF and purified. Cells from 3 to 4 women were grouped and total mRNA was subjected to qPCR with DCN and b-actin primers. DCN was quantified by qPCR and normalized to β-actin expression. Data represent the mean ± SEM of three independent experiments. Gene expression was calculated as fold change relative to CGC that was set to 1. (P = 0.02). b. DCN mRNA expression in MGCs obtained from different stages of follicular development. MGCs were collected during IVM from small antral follicles (< 14 mm) and during IVF procedures from preovulatory follicles (> 17 mm). Data represent the mean ± SEM of three independent experiments. Gene expression was calculated as fold change relative to IVM that was set to 1. (P = 0.001). c. DCN mRNA expression in CGCs according to maturation stage of the related oocyte. CGCs were collected from cumulus-oocyte complex retrieved from women undergoing in-vitro fertilization (IVF) procedure. Cells were grouped according to the maturation stage of the related oocyte: GV, MI, and MII. Extracted mRNA was subjected to qPCR with DCN primers and b-actin primers. Data represent the mean ± SEM of three independent experiments. Gene expression was calculated as fold change relative to GV that was set to 1. (P < 0.05). d. Characterization of the optimal incubation time for hCG effect on DCN mRNA expression in MGCs cell culture. MGCs were collected during IVF procedure and cultured. hCG (1 or 10 unit/ml) was added to the medium after 0–6 incubation days with a daily medium exchange. Cells were collected 24 h after hCG stimulation and the total mRNA was subjected to qPCR with DCN primer and b-actin primers. Data represent the mean ± SEM of three independent experiments. Gene expression was calculated as fold change relative to the control sample that was set to 1. Different letters represent significant results. (P < 0.05) (e) The conditioned media of the experiment in d was collected for DCN concentration measurement using EIA kit. Concentrations are in absolute values. * represent significant differences (P < 0.05). The results are expressed as mean ± SEM of three independent experiments. f The protein level of DCN was determined by western blotting in three independent experiments. Actin was used as a control. gDCN mRNA expression in MGCs cell culture during 24 h. MGCs were aspirated from preovulatory follicles (> 17 mm) during IVF and cultured for 4 days with a daily medium exchange, then, 10 unit/ml of hCG was added to the medium. Cells were collected at starting point and at 2 h, 4 h, 6 h, 8 h, and 24 h. Data represent the mean ± SEM of three independent experiments. Gene expression was calculated as fold change relative to T0 sample that was set to 1. (P DCN = 0.003). * marks the significant result

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