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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: Mol Cell Endocrinol. 2014 Aug 1;395(0):19–31. doi: 10.1016/j.mce.2014.07.020

Fig. 2.

Fig. 2

BCL6 and STAT5 show reciprocal occupancy at transcription start sites of Socs2, Cish, and Bcl6. (A) Peaks of BCL6 occupancy determined by ChIP-Seq were identified by HPeak at the transcription start sites (TSS) of Socs2, Cish and Bcl6. Shown are the UCSC Genome Browser profiles for BCL6 occupancy in 3T3-F442A adipocytes treated with GH for 0 or 48 h. Y axis scales were set at 60 for Socs2, 35 for Cish, and 20 for Bcl6 as determined by maximum peak height in the absence of GH as calculated by HPeak. Top panel: Socs2. TSS: mouse chromosome (chr) 10: 94879491. Peak location 0 h GH: chr10: 94879276-94880050; 48 h GH: chr10: 94879326-94879975. Predicted BCL6/STAT consensus sites: chr10: 94879539-94879547 and chr10: 94879550-94879558. Second panel: Cish. TSS: chr9: 107199020. Peak location 0 h GH: chr9: 107198726-107199400; 48 h GH: chr9: 107198751-107199150. Predicted BCL6/STAT consensus sites: chr9: 107198991-107198999 and chr9: 107199002-107199010. Third panel: Bcl6. TSS: chr16: 23988698. Peak location 0 h GH: chr16: 23988401-23988850; 48 h GH: chr16: 23988776-23988900. Predicted BCL6/STAT consensus sites: chr16: 23988676-23988684 and chr16: 23988717-23988725. Bottom panel: Peak profile at Bcl6 promoter region with expanded scale to visualize peaks in detail. Arrows to the left of gene names indicate direction of transcription, from 5′ to 3′. Gene structure diagrammed at bottom of each profile. (B) 3T3-F442A adipocytes were treated with GH for 0 or 48 h and ChIP was carried out using antibodies against BCL6 and STAT5. ChIP DNA was probed with primers specific to the BCL6 occupancy sites corresponding to ChIP-Seq (peaks) on the Socs2 (lanes 1 and 2), Cish (lanes 3 and 4) and Bcl6 (lanes 5 and 6) genes. (C) 3T3-F442A adipocytes were treated with GH for 0, 4 or 48 h. RNA was prepared and analyzed by qPCR using primers for Socs2, Cish and Bcl6. Bars show mean + SE of triplicates in an experiment which was repeated at least 3 times.