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. 2013 Apr 11;8(4):e61055. doi: 10.1371/journal.pone.0061055

Figure 5. Chromatin association of CTR9 requires kinase activity inhibited by AG490.

Figure 5

A, Cells were transfected with control siRNA or CTR9 siRNA for 48 hrs and treated with DMSO (mock) or AG490 (100 uM) for 4 hrs prior to IL-6 plus IL-6sR (20 ng/ml each) stimulation for 30 min. The c-Fos mRNA induction level was measured by RT-qPCR. B, Cells were treated as in A and a ChIP assay was performed with an anti-CTR9 antibody. Bound DNAs were analyzed by quantitative PCR using primers specific to c-Fos genomic loci. C, Cells were treated with DMSO (mock), 100 uM or 200 uM of AG490 for 12 hrs prior to IL-6 plus IL-6sR (20 ng/ml each) stimulation for 30 min. Cells were fractionated into chromatin -unbound and -bound fractions and blotted with anti-CTR9, anti-STAT3, anti-TBP, and anti-GAPDH antibodies. D, Negative function of PAFc in IL-6-responsive gene regulation. At TSS of IEG loci in basal condition, PAFc is associated with NELF. Upon IL-6 stimulation, PAFc along with NELF dissociates from the elongation block to allow transcriptional elongation to occur. SIE: Serum Inducible Element (STAT3 binding site). n.s. = not significant, **p<0.01 by Student's t test. Error bars represents SD (n = 3).