Fig. 2

Effects of neuregulin-1 (NRG1) knockdown on chemokines and cytokines secretion 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 media were collected at 48 h. Concentrations of chemokines and cytokines were measured and analyzed in media using Bio-Plex-Pro-Rat Cytokine, Chemokine, and Growth Factor Magnetic Bead-Based Assays (Bio-Rad, USA) coupled with the Luminex-200 system. Cytokine, Chemokine, and Growth Factors are expressed in picogram per ml per milligram (pg/ml/mg) cellular protein. 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. Abbreviations- IL: Interleukin, G-CSF: granulocyte colony-stimulating factor, GM-CSF: granulocyte-macrophage colony-stimulating factor, GRO-KC: Keratinocyte chemoattractant (KC)/human growth-regulated oncogene (GRO), IFNγ: Interferon-gamma, MCSF: Macrophage-colony stimulating factor, MCP1: Monocyte chemoattractant protein-1, MIP-3α: Macrophage Inflammatory Protein 3α, TNFα: tumor necrosis factor α, VEGF: vascular permeability factor/vascular endothelial growth factor, RANTES: Regulated upon Activation, Normal T Cell Expressed and Presumably Secreted.