Fig. 3

Effects of neuregulin-1 (NRG1) knockdown on kinases in granulosa cells (GCs). Pregnant Mare Serum Gonadotropin (PMSG) primed GCs were isolated from two ovaries per rat per sample and grown in culture dishes. GCs were transiently transfected with small-interfering-NRG1 (siNRG1) RNA and scramble-green-fluorescent-protein (GFP) RNA (negative control). Thereafter, cells were maintained in serum-free media and harvested at 24 and 48 h. Representative immunoblots showing phospho(p)- and total phosphoinositide 3-kinase (PI3K), serine/threonine-protein kinase (AKT/Protein Kinase B) and extracellular signal-regulated kinases (ERK1/2) protein expressions in GCs. Equal amounts of protein were applied to each lane. β-actin was used as an internal control. Bar diagrams represent the densitometric analyses of immunoblots. All bar graphs represent the mean ± SEM of results from three individual experiments (n = 3). Asterisks (*) represent unpaired Student t-test, *p ≤ 0.01, **p ≤ 0.001, ***p ≤ 0.0001, NS-not significant