Fig. 2
CFDTW reduces the expression of PKP3 by augmenting the PKP3 promoter methylation in PCOS rats. Normal rats served as controls and PCOS rats were further treated with CFDTW. A, Rat estrous cycle (P: proestrus; E: estrus; M: post-estrus; D: during estrus). B, Rat ovarian weight. C, Serum levels of T in rats. D, Serum levels of E2 in rats. E, Serum levels of LH in rats. F, Serum levels of FSH in rats. G, HE staining images of rat ovarian tissues, scale bar 50 μm. H, Schematic diagram of CpG islands in the rat PKP3 gene promoter. I, The methylation degree of the PKP3 promoter in rat ovarian tissues determined by MSP. J, Methylation status of CpG islands in the PKP3 promoter in rat ovarian tissues determined by BSP. The color of the circle at each CpG site represents the percentage of methylation. K-L, RT-qPCR detection (K) and immunohistochemistry analysis (L) of PKP3 expression in the rat ovarian tissue. M, Representative microscopic views of the ovarian granulosa cells isolated from rats under an inverted microscope, scale bar 50 μm. N, Immunofluorescence staining analysis of FSHR expression (green fluorescence) in the ovarian granulosa cells isolated from rats, scale bar 25 μm. O, The methylation degree of the PKP3 promoter in rat ovarian granulosa cells determined by MSP. P, Methylation status of CpG islands in the PKP3 promoter in rat ovarian granulosa cells determined by BSP. The color of the circle at each CpG site represents the percentage of methylation. Q, RT-qPCR detection of PKP3 expression in rat ovarian granulosa cells. * p < 0.05 vs. normal rats or control cells. # p < 0.05 vs. PCOS rats or those treated with CFDTW. n = 8 for rats following each treatment