体外研究：

样本类型（Sample type）：Semen Preparation

应用目的（Purpose）：Using CMA3 staining to determine quality chromatin packaging

实验方法（Assay）：CMA3 staining thin smears were prepared, air dried, andfixed in methanol/acetic acid 3:1 at room temperature for 20 min. The slides were air-dried and stained with 60–100µl CMA3 in a dark chamber for 20 min. Slides were then washed in McIlvaine buffer and mounted using Dabco. Two hundred spermatozoa were evaluated under a Nikonfluorescent microscope.

体外研究：

样本类型（Sample type）：Semen aliquots

应用目的（Purpose）：Using CMA3 staining to assess the packaging quality of the chromatin in sperm

实验方法（Assay）：The spermatozoa were washed again, fixed in methanol/glacial acetic acid (3:1) at 4℃ for 5 min and then spread on slides. For CMA3 staining, each slide was treated for 20 min with 100µl of CMA3 solution (0.25 mg/ml in McIlvaine buffer, pH 7.0, containing 10 mM MgCl2). The slides were then rinsed in buffer and mounted with buffered glycerol. Microscopic analysis of the slides was performed on a fluorescent Zeiss axioplane microscope with the appropriate filters.

体外研究：

细胞类型（Cell type）：PC3 cells

药物配制（Preparation）：Chromomycin A3 was dissolved in ethanol as 1:1000 stock solution.

实验方法（Assay）：PC3 cells were seeded at indicated densities in 12-well plates in triplicates and treated with DMSO, atorvastatin (10 μM), cyclohexamide (10 μg/ml) and chromomycin A3 (5 μM) or atorvastatin plus cyclohexamide or chromomycin A3. The cells were cultured for 2, 3 and 4 d, trypsinized, and counted by the cell counter Cellometer Auto T4.