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. Author manuscript; available in PMC: 2012 Mar 15.
Published in final edited form as: Oncogene. 2011 Apr 4;30(37):3943–3952. doi: 10.1038/onc.2011.105

Fig. 5. EGF increases occupancies of Brf1 and H3S28ph, but decreases H3K27me3 in tRNALeu and 5S rRNA promoters.

Fig. 5

(A) EGF-mediated occupancy in the tRNALeu promoter. JB6 cells were treated with or without EGF and ChIP assays were performed using Brf1, H3S28ph, H3K27me3 and H3 antibodies and qPCR with specific tRNALeu primers to quantify the amplified DNA. (B) EGF enhances Brf1, H3S28ph occupancy in the 5S rRNA promoter. ChIP assays were performed as indicated in Fig. 4 and qPCR was used with 5S rRNA primers to quantify the DNA. (C) Expression of mutant H3S28A reduces H3S28ph occupancy in tRNALeu and 5S rRNA promoters. These cells were treated as designed Fig 4B and C. Chromatin was extracted from these cells to perform ChIP assays with H3S28ph antibody. The relative occupancy of the proteins was calculated based on the control (no EGF treatment). All values shown are the means ± SEM of at least three independent chromatin preparations.