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Percentage of cultured cell-populations that stained positively and/or negatively for apoptotic markers cleaved caspase-3 and cleaved PARP, following DNA damage treatments induced by doxorubicin and TNF-alpha co-treatments

机译:在多柔比星和TNF-α共处理诱导的DNa损伤处理后,凋亡标记阳性和/或阴性染色的培养细胞群的百分比切割caspase-3和切割的paRp

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摘要

The present dataset contains the source data for Figure 2B of Tentner et al. (2012). The data shows the percentage of cultured cell-populations that stained positively and/or negatively for apoptotic markers cleaved caspase-3 and cleaved PARP, following DNA damage treatments induced by various doses of doxorubicin (0, 2 and 10 µmole/L) in the presence (100 ng/mL) or absence (0 ng/mL) of TNF-alpha co-treatment. For the six treatment conditions investigated, cell counts were made by flow cytometry at times 6, 12, 24, and 48 h following treatment; CULTURE DETAILS: U2OS cells were obtained from ATCC were maintained at 21% oxygen and 5% CO2 in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum, penicillin, streptomycin, 2mM L-glutamine, and used within 15-20 passages. The first thymidine block was released by washing the plates three times with PBS, and incubating them in fresh thymidine-free media for 12 h. A second thymidine block was then performed by re-addition of thymidine to 2.5 mM followed by incubation for an additional 18 h. Media was aspirated, plates were washed 3 with PBS, and replaced with fresh media in the presence or absence of 10 mM aphidicolin; ANALYSIS DETAILS: See supplementary journal publication; RESULT: The authors of the supplementary journal publication conclude that TNF enhances dose-dependent cell death following doxorubicin-induced DNA damage with minimal affect on dose-dependent cell-cycle arrest.
机译:本数据集包含Tentner等人的图2B的源数据。 (2012)。数据显示,在不同剂量的阿霉素(0、2和10 µmole / L)诱导的DNA损伤处理后,凋亡标记切割的caspase-3和切割的PARP阳性和/或阴性染色的培养细胞群体的百分比。 TNF-α联合治疗的存在(100 ng / mL)或不存在(0 ng / mL)。对于所研究的六个治疗条件,在治疗后的第6、12、24和48小时通过流式细胞仪进行细胞计数。文化细节:从ATCC获得的U2OS细胞在补充有10%胎牛血清,青霉素,链霉素,2mM L-谷氨酰胺的Dulbecco改良Eagle培养基中维持21%的氧气和5%的CO2的使用,并在15至20代中使用。通过用PBS洗涤板3次并将其在无胸腺嘧啶的新鲜培养基中孵育12小时,释放第一个胸腺嘧啶核苷。然后通过将胸苷重新添加至2.5 mM进行第二个胸苷封闭,然后再孵育18小时。吸出培养基,将板用PBS洗涤3次,并在存在或不存在10 mM蚜虫碱的情况下用新鲜培养基替换;分析详细信息:请参阅期刊补充出版物;结果:该补充刊物的作者得出结论,TNF增加了阿霉素诱导的DNA损伤后剂量依赖性细胞死亡,而对剂量依赖性细胞周期停滞的影响最小。

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