PQM-1 controls hypoxic survival via regulation of lipid metabolism.
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Worms were exposed to 5 mM CoCl2 at the early day 1 of adulthood stage for 6 hr and 20 hr. Untreated worms were used as controls. Worms were collected, RNA isolated and hybridized on 4x44K C. elegans arrays (Agilent) at 60C overnight, as previously described 1). Three biological replicates were used. Significant differentially-expressed gene sets were identified using one or two-class SAM 2). 1) Shaw, W. M., Luo, S., Landis, J., Ashraf, J. & Murphy, C. T. The C. elegans TGF-beta Dauer pathway regulates longevity via insulin signaling. Curr Biol 17, 1635-1645, doi:10.1016/j.cub.2007.08.058 (2007). 2) Tusher, V. G., Tibshirani, R. & Chu, G. Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 98, 5116-5121, doi:10.1073/pnas.091062498 (2001).
本实验将秀丽隐杆线虫（C. elegans）于成虫早期第1天置于5 mM氯化钴（CoCl₂）中分别处理6小时与20小时，以未处理的线虫作为空白对照组。收集各组线虫并提取总RNA，于60℃条件下过夜杂交至4×44K规格的秀丽隐杆线虫基因芯片（安捷伦，Agilent），实验流程参照此前发表的文献1。本实验设置三次生物学重复，采用单分类或双分类显著性微阵列分析（SAM，Significance Analysis of Microarrays）方法筛选得到显著差异表达基因集。 1) Shaw, W. M., Luo, S., Landis, J., Ashraf, J. & Murphy, C. T. The C. elegans TGF-beta Dauer pathway regulates longevity via insulin signaling. Curr Biol 17, 1635-1645, doi:10.1016/j.cub.2007.08.058 (2007). 2) Tusher, V. G., Tibshirani, R. & Chu, G. Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 98, 5116-5121, doi:10.1073/pnas.091062498 (2001).
