Figure 5.

Nuclear run-on assays of nascent ³²P-labeled RNA transcripts isolated from the nuclei of FTO-2B cells treated with arachidonate in the absence or presence of T0901317. Cells were set up for experiments as described in Materials and Methods. After incubation for 6 h in medium B with or without 100 μM sodium arachidonate (A) in the absence or presence of 1 μM T0901317 (T) as indicated, the cells from four 150-mm dishes were pooled, lysed, and subjected to nuclear run-on assay as described in Materials and Methods. After hybridization with the indicated cDNA probe, the washed filters were exposed to Kodak X-Omat film with a Bio-Max (Kodak) intensifying screen at −80°C for 24 h. Relative levels of the transcripts for each gene (shown at right) were quantified with a Fuji Bio-Imaging analyzer. A blank value corresponding to the vector band was subtracted from all values.