Fig. 4

The effects of the GPER-specific antagonist (G-15) and/or bisphenol A (BPA) and bisphenol S (BPS) on gene expression of ovine granulosa cells (GC), according to changes over time. The expression of eight genes (CHAC1, JUN, LSMEM1, NR4A1, TXNIP, ARRDC4, KLF9 and LIF) were assessed in ovine GC. Gene expression was determined after 12, 24 and 48 h of culture in complemented serum-free McCoy’s 5A media in the presence or absence (control) of BPA or BPS at 50 μM. Total messenger RNA (mRNA) was extracted and reverse transcribed from ovine GC, then real-time polymerase chain reaction (qPCR) was performed. To normalise gene expression, the geometric mean of two housekeeping genes (β-actin [ACTB] and ribosomal protein L19 [RPL19]) was used. The results are expressed as the mean ± standard error of the mean of eight independent cultures and normalised to the mean of the control condition at 12 h. Statistical analysis was performed between the conditions of a same time (12, 24 or 48 h). Bars with different superscripts indicate a significant difference (p ≤ 0.05)