Rat GRP78 mRNA, rno-miR-144, rno-miR-376a, and rno-miR-451 expression in primary rat granulosa cells induced by FSH and hCG.

<p>Primary rat granulosa cells were prepared, and the indicated reagents were added to the medium after 24 h of culture. Cells were then incubated with FSH (30 ng/mL) and estradiol (10 nM) for 48 h. Subsequently, hCG (30 ng/mL) was added to the culture medium, as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108997#s2" target="_blank">Materials and Methods</a>. Total RNA was isolated, and GRP78 mRNA (A), rno-miR-144 (B), rno-miR-376a (C), and rno-miR-451 (D) expression levels were measured using real-time RT-PCR as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108997#s2" target="_blank">Materials and Methods</a>. The amounts of GRP78 mRNA, rno-miR-144, rno-miR-376a, and rno-miR-451 in the hCG 0 h group were set at 1. Data were normalized for 18S rRNA (for GRP78 mRNA) and 4.5S RNA(H) (for rno-miR-144, rno-miR-376a, and rno-miR-451) levels in each sample and represent the mean ±SE of 3 independent experiments. *, significantly different from the control value at hCG 0 h, <i>P</i><0.05.