Effects of RU486 on the dorsal skin of stressed mice.

A: Photographs of mice back skin on day 2 and day 13 after epilation showing that the mice received RU486 injection had a significantly darker observable skin color than CUMS mice. B: MITF expression was analyzed by immunoblotting. C: TYR expression was analyzed by immunoblotting. D: POMC expression was analyzed by immunoblotting. β-Actin expression was indicated as a loading control. Western blot assay are representative of three experiments. Densitometric scanning of band intensities obtained from three separate experiments was used to quantify change of proteins expression. Three animals were used for each data point. Data are showed in mean ± SEM. E: The mRNA expression levels of UCN1, POMC, and CYP11A1 in mouse skin. The expression levels of each gene were normalized against β-Actin then calculated as fold change using the comparative 2-ΔΔCT method. Data are showed in mean ± SEM, n = 8. The data were analyzed by one-way ANOVA with Tukey's post hoc test. * P<0.05, *** P<0.001, compared with control; &P<0.05, &&P<0.01, &&&P<0.001, compared with CUMS.

A: 脱毛后第2天和第13天的小鼠背部皮肤照片，结果显示注射RU486的小鼠皮肤肉眼可见颜色显著深于CUMS（慢性不可预知温和应激，Chronic Unpredictable Mild Stress）组小鼠。B: 采用免疫印迹法（immunoblotting）分析MITF (Microphthalmia-associated transcription factor，小眼畸形相关转录因子)的蛋白表达水平。C: 采用免疫印迹法分析TYR (Tyrosinase，酪氨酸酶)的蛋白表达水平。D: 采用免疫印迹法分析POMC (Pro-opiomelanocortin，阿黑皮素原)的蛋白表达水平；以β-肌动蛋白(β-Actin)作为上样内参。本次蛋白质免疫印迹实验（Western blot）结果为3次独立重复实验的代表性结果。通过对3次独立实验得到的条带强度进行光密度扫描，以量化蛋白表达的变化情况。每个数据点均使用3只小鼠进行实验。实验数据以均值±标准误表示。E: 小鼠皮肤中UCN1 (Urocortin 1，尿皮质素1)、POMC及CYP11A1 (Cytochrome P450 11A1，细胞色素P450家族11亚家族A成员1)的mRNA表达水平。各基因的表达水平均以β-肌动蛋白作为内参进行归一化处理，随后采用相对定量2-ΔΔCT法计算其表达倍数变化。实验数据以均值±标准误表示，样本量n=8。数据采用单因素方差分析（one-way ANOVA）结合Tukey事后检验进行统计学分析。与对照组相比，* P<0.05，*** P<0.001；与CUMS组相比，& P<0.05，&& P<0.01，&&& P<0.001。