Fig 1: Effects of MuV on testosterone synthesis.(A) Expression of steroidogenic enzymes. Leydig cells of C57BL/6 mice were infected with 5 MOI MuV for specific durations. The uninfected cells served as controls (Ctrl). Total RNAs were extracted and the relative mRNA levels of 3ß-hydroxysteroid dehydrogenase (3ß-HSD), cytochrome P450 side-chain cleavage enzyme (P450scc), and steroidogenic acute regulatory protein (StAR) were determined with real-time qRT-PCR by normalizing to ß-actin. (B) Protein levels of the enzymes. Leydig cells were infected as described in (A). The protein levels of 3ß-HSD, P450scc and StAR were determined with Western blot using specific antibodies. (C) Testosterone level. Leydig cells were treated as described in (A). The testosterone level in the culture medium was measured with ELISA. (D) Testosterone synthesis in vivo. One testis of five-week-old C57BL/6 mice were injected with 1 × 107 PFU MuV in 10 µl of PBS. The contralateral testis was injected with an equal volume of PBS alone for the Ctrl. At the indicated time points post injection, the testes were lysed and the testosterone level were measured with ELISA. Each dot indicates the testosterone level of individual testes. Data are means ± SEM (n = 5 mice). **p < 0.01.
Fig 2: The effect of chemerin on StAR and P450SCC protein abundance. The influence of chemerin (100, 200 ng/mL) on the protein abundance of steroidogenic acute regulatory protein (StAR; (a)) and P450 side-chain cleavage enzyme (P450SCC; (b)) in the in-vitro-incubated endometrial tissue explants collected from pigs on days 10 to 11, 12 to 13, 15 to 16, and 27 to 28 of pregnancy, and on days 10 to 11 of the oestrous cycle. The protein abundance of StAR and P450SCC was evaluated using Western blot analysis. Upper panels—representative immunoblots, lower panels—densitometry analysis of target proteins’ relative content normalised with the actin protein. Data are presented as the mean ± standard error of the mean (n = 5). Bars with different letters are significantly different at p < 0.05. C—control; CH100—chemerin 100 ng/mL; CH200—chemerin 200 ng/mL.
Fig 3: Measurement of protein levels of steroidogenic acute regulatory protein (STAR), CYP11A1, and CYP17A1 in the testes of lipopolysaccharide (LPS; 1 mg/kg)-injected mice fed ginseng berry extract (GBE; 50 and 100 mg/kg/day). Western blotting analysis was performed with eight mice testes from each experimental groups (A) and total protein was normalized to the expression of ß-ACTIN (B). *p<0.05, **p<0.01, ***p<0.001.
Fig 4: Measurement of protein levels of steroidogenic acute regulatory protein (STAR), CYP11A1, and CYP17A1. TM3 cells were treated with 0, 1, 10, 25, and 50 µg/mL of ginseng berry extract (GBE) (A) and 0, 0.5, 1, 5, 10, and 50 ng/mL of lipopolysaccharide (LPS) (C). Western blotting was performed with the gene specific antibodies for STAR, CYP11A1, CYP17A1, and ß-ACTIN. Total protein was normalized with the expression of the ß-ACTIN gene (B and D). *p<0.05, **p<0.01, ***p<0.001.
Fig 5: Measurement of mRNA levels of steroidogenic acute regulatory protein (STAR), CYP11a1, and CYP17a1 in mice testes. Mice were fed with 50 mg/kg/day of ginseng berry extract (GBE), with or without sleep deprivation for two days. Each group had eight mice. *p<0.05, **p<0.01.
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