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. 2018 Feb 8;13(2):e0192458. doi: 10.1371/journal.pone.0192458

Fig 5. Roles of AREG and fibronectin in progesterone production of granulosa cells.

Fig 5

(A): Progesterone concentrations in culture supernatants of granulosa cells treated for 24 h with or without AREG on uncoated or fibronectin-coated wells. Values are the mean ± SEM of three replicates. Data were statistically analyzed by two-way ANOVA. Because the significant difference was observed for each factor, the comparison analysis was performed by the Student’s t-test. *; AREG significantly increased the area of granulosa cells cultured on fibronectin-coated wells or free wells (p<0.05). Free: granulosa cells cultured on uncoated wells; Fibronectin: granulosa cells cultured on fibronectin-coated wells; C (control): granulosa cells treated without AREG; AREG: granulosa cells treated with AREG. (B): Expression of genes involved in progesterone production (Star, Cyp11a1, and Hsd3b1) in granulosa cells treated for 24 h with or without AREG on uncoated or fibronectin-coated wells. Values are the mean ± SEM of three replicates. Data were statistically analyzed by two-way ANOVA. Because the significant difference was observed for each factor, the comparison analysis was performed by the Student’s t-test. *; AREG significantly increased the area of granulosa cells cultured on fibronectin-coated wells or free wells (p<0.05). **AREG significantly increased the area of granulosa cells cultured on fibronectin-coated wells (p<0.05). Free: granulosa cells cultured on uncoated wells; Fibronectin: granulosa cells cultured on fibronectin-coated wells; C (control): granulosa cells treated without AREG; AREG: granulosa cells treated with AREG. (C): Additional effects of Y15 on AREG-induced progesterone production in granulosa cells cultured on fibronectin-coated wells or free-wells. Values are the mean ± SEM of three replicates. *; Y15 treatment significantly decreased the progesterone level when granulosa cells were cultured on fibronectin-coated wells (p<0.05). (D): The treatment with Y15 inhibited AREG-induced expression of genes involved in progesterone production (Star, Cyp11a1, and Hsd3b1) in granulosa cells. Values are the mean ± SEM of three replicates. *; Y15 treatment significantly inhibited the gene expression levels compared with each control (p<0.05).