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. Author manuscript; available in PMC: 2015 Mar 25.
Published in final edited form as: Exp Hematol. 2010 May 5;38(8):629–640.e1. doi: 10.1016/j.exphem.2010.04.004

Fig. 2. PU.1 binds to conserved sequences in the 23a cluster promoter.

Fig. 2

A) Alignment of conserved regions of the miR-23a cluster promoter from mouse and human. Numbering is relative to human start site. Putative PU.1 binding sites (AGGAA core sequence) are boxed in red. B) EMSA demonstrating PU.1 binds in vitro to each of the 4 conserved PU.1 sites (blue underlined sequence used as probes). Nuclear extracts (NE) prepared from mock (ctrl), or PU.1 transfected 293T cells. Controls include incubations with unlabeled wildtype and mutant competitor oligos as well as antibody to PU.1. C) Chromatin immunoprecipitation analysis. Sheared chromatin extracts were prepared from PUER cells untreated or treated with OHT for 7 days. Chromatin was immunoprecipitated with anti-PU.1 or as a negative control anti-GATA1 antibody. Quantitative SYBR green PCR was performed to determine whether the 23a promoter was present in the immunoprecipitates. Fold-enrichment of the 23a promoter above what was detected in GATA1 precipitates from untreated PUER cells is shown.