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. 2022 Jul 26;15:86. doi: 10.1186/s13048-022-01021-0

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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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.