Temporal analysis of RNA turnover in Interferon Gamma treated bone marrow-derived macrophages
收藏资源简介:
Bone Marrow derived macrophages (BMDM) were grown using L929 conditioned medium. On day 7, BMDM were mock-treated (16x 15cm dishes) or treated (16x 15cm dishes) with 10U/ml interferon gamma at time zero. 4-thiouridine (4sU) was then used to label newly transcribed RNA every 30 minutes for 8h (0 to 30, 30 to 60 etc. to give 16 discrete successive labellings) in mock and interferon gamma-treated samples. 16x Mock and 16x treated total samples were then isolated (32 in total) and newly transcribed RNA isolated from each total RNA sample using a protocol described in Dolken et al (PMID: 18658122). For analysis, 32 (16x mock and 16x treated) newly-transcribed RNA samples (every 30 minutes for complete 8h time course) and 16 (8x mock and 8x treated) total RNA samples (every hour for 8h time course) were hybridized to Affymetrix GeneST v1.0 arrays.



