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.

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.