Fig 1: Plasma concentrations of RARRES2 (A), ADIPOQ (B), testosterone (C), and estradiol (D) measured by ELISA assay. Results were mean ± SEM of eight independent determinations. Statistically significant differences (p < 0.05) between points in each graph are indicated by different letters (a < b < c), identical letters indicate a lack of significant differences. ADIPOQ, adiponectin. C, control. ELISA, enzyme-linked immunosorbent assay. PCOS, polycystic ovarian syndrome. RARRES2, retinoic acid 109 receptor responder protein 2. SEM, standard error of the mean. VD3, vitamin D3.
Fig 2: Representative micrographs of immunohistochemical RARRES2 and CMKLR1 localization in uterus obtained from control ((A) and (F), respectively), VD3-treated ((B) and (G), respectively), induced PCOS ((C) and (H), respectively), and VD3-treated PCOS ((D) and (I), respectively) rats. Arrows indicate positive RARRES2 and CMKLR1 staining within LE and GE in all examined groups ((A–D) and (F–I), respectively). Asterisks indicate positive CMKLR1 immunoreaction in stroma cells (F–I). Control sections ((E) and (J), respectively) showed no positive staining. Charts represent the intensity of RARRES2 (K) and CMKLR1 (L) immunostaining expressed as the relative optical density (ROD) in LE, GE, and stroma. Statistically significant differences (p < 0.05) between points in each graph are indicated by different letters (a < b < c), identical letters indicate a lack of significant differences. Scale bar 50 µm. C, control. CMKLR1, chemerin chemokine-like receptor 1. GE, glandular epithelium. LE, luminal epithelium. RARRES2, retinoic acid 109 receptor responder protein 2. PCOS, polycystic ovarian syndrome. SEM, standard error of the mean. VD3, vitamin D3.
Fig 3: mRNA and protein expression of RARRES2 (A,B), CCRL2 (C,D), CMKLR1 (E,F), and GPR1 (G,H) in rat uterus. mRNA expression was determined using real-time PCR and presented relative to Gapdh; the protein expression was detected via Western blot assay, each protein abundance was evaluated densitometrically and expressed as the ratio relative to the ACTB abundance. Results were mean ± SEM of eight independent determinations. Statistically significant differences (p < 0.05) between points in each graph are indicated by different letters (a < b < c < d), identical letters indicate a lack of significant differences. ACTB, ß-actin. C, control. CCRL2, C-C motif chemokine receptor-like 2. CMLR1, chemerin chemokine-like receptor 1. Gapdh, glyceraldehyde-3-phosphate dehydrogenase. GPR1, G protein-coupled receptor 1. PCR, polymerase chain reaction. PCOS, polycystic ovarian syndrome. RARRES2, retinoic acid 109 receptor responder protein 2. SEM, standard error of the mean. VD3, vitamin D3.
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